Endurance Exercise Mobilizes Developmentally Early Stem Cells into Peripheral Blood and Increases...

Research ArticleEndurance Exercise Mobilizes Developmentally EarlyStem Cells into Peripheral Blood and Increases Their Number inBone Marrow Implications for Tissue Regeneration

Krzysztof Marycz12 Katarzyna Mierzejewska3 Agnieszka Umieszek12 Ewa Suszynska3

Iwona Malicka4 Magda Kucia35 and Mariusz Z Ratajczak35

1Faculty of Biology University of Environmental and Life Sciences 50-375 Wroclaw Poland2Wroclaw Research Center EIT+ 54-066 Wroclaw Poland3Department of Regenerative Medicine Medical University of Warsaw 02-091 Warsaw Poland4Department of Physiotherapy University School of Physical Education 51-617 Wroclaw Poland5Stem Cell Biology Program James Graham Brown Cancer Center Louisville KY 40202 USA

Correspondence should be addressed to Krzysztof Marycz krzysztofmaryczinteriapland Mariusz Z Ratajczak mariuszratajczaklouisvilleedu

Received 18 February 2015 Accepted 26 March 2015

Academic Editor Irma Virant-Klun

Copyright copy 2015 Krzysztof Marycz et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Endurance exercise has been reported to increase the number of circulating hematopoietic stemprogenitor cells (HSPCs) inperipheral blood (PB) as well as in bonemarrow (BM)We therefore became interested in whether endurance exercise has the sameeffect on very small embryonic-like stem cells (VSELs) which have been described as a population of developmentally early stemcells residing in BMMice were run daily for 1 hour on a treadmill for periods of 5 days or 5 weeks Human volunteers had trained inlong-distance running for one year six times per week FACS-based analyses and RT-PCR ofmurine and humanVSELs andHSPCsfrom collected bone marrow and peripheral blood were performed We observed that endurance exercise increased the number ofVSELs circulating in PB and residing in BM In parallel we observed an increase in the number of HSPCsThese observations weresubsequently confirmed in young athletes who showed an increase in circulatingVSELs andHSPCs after intensive running exerciseWe provide for the first time evidence that endurance exercisemay have beneficial effects on the expansion of developmentally earlystem cellsWe hypothesize that these circulating stem cells are involved in repairingminor exercise-related tissue and organ injuries

1 Introduction

Bonemarrow (BM) contains a variety of stem cells includinghematopoietic stemprogenitor cells (HSPCs) endothelialprogenitor cells (EPCs)mesenchymal stem cells (MSCs) andthe dormant population of stem cells from early embryonicdevelopment that have been named very small embryonic-like stem cells (VSELs) [1ndash3] It has been shown that BM-derived stem cells especially HSPCs circulate in peripheralblood (PB) at a very low level under steady-state conditions[4] This circulation allows the pool of stem cells maintainedin BM to be spread to bones located in distant parts ofthe body Another important suggested purpose of this

circulation is that various types of such circulating stemcells play a role in ldquopatrollingrdquo peripheral tissues to preventinfections and tissue damage [5]

Evidence has also accumulated that HSPCs and EPCsexpand in bonemarrow (BM) in response to endurance exer-cise and are subsequently mobilized into peripheral blood(PB) [6ndash9] Therefore we became interested in whether thepool of BM-residingVSELswould respond in a similar way asHSPCs to endurance exerciseThese cells as demonstrated inseveral reports have the ability to differentiate into cells fromall three germ layers [10] and play an important role in tissueand organ rejuvenation [11] and their number positivelycorrelates with life span in experimental animals [12]

Hindawi Publishing CorporationStem Cells InternationalArticle ID 395971

2 Stem Cells International

Egress of stem cells from the BM is triggered by activa-tion of the complement cascade which releases importantactive cleavage fragments such as the C5 component C5athat induce granulocytes and monocytes in BM to releaseproteolytic enzymes that attenuate stem cell retention signalsin BM niches and permeabilize the BMndashPB barrier thusfacilitating the egress of stem cells [13] The major chemoat-tractant for stem cells in PB is sphingosine-1-phosphate (S1P)and in the process of mobilization certain other factors arealso involved including 120572-chemokine stromal-derived factor1 (SDF-1) [14] It has also been proposed that mobilizationof particular types of stem cells may be modulated by stemcell type-specific factors such as vascular endothelial growthfactor (VEGF) which is involved in mobilization of EPCs[15] Our recent research demonstrated that mobilization ofstem cells not only is triggered by but also correlates withactivation of the complement cascade and its activation canbe easilymeasured in PB using commercially available ELISAassays to detect for example the C5b-C9 membrane attackcomplex (MAC) [16]

Our data confirm that HSPCs are released from BMinto PB in response to physical exercise and demonstratefor the first time that in parallel VSELs are also mobilizedImportantly we demonstrate a positive effect of exercise onexpansion of this primitive pool of stem cells in BM Sincethese small cells may differentiate into several types of cellsacross the three germ layers their increase may explain ina novel way the positive effect of regular exercise on tissueand organ rejuvenation Of note a similar positive effecton this pool of cells by caloric restriction has recently beendemonstrated [17]Therefore VSELsmay be able to reconcilereported observations of a positive effect of both exercise andcaloric restriction on life quality and extension of life spanHowever we are aware that more direct data are necessary tofully support this novel concept

2 Materials and Methods

21 Animal Care and Use Conducted studies were approvedby the II Local Ethical Committee Environmental and LifeScience University and adhered to American College ofSports Medicine (ACSM) animal care standards

22 Experimental Animals Theexperiments were performedon 90 4-week-old C57BL6 mice Sedentary control (SED)and exercise-trained (EX) C57Bl6 mice were housed threeper cage in an ultraclean facility on ventilated racks and wereprovided food and water ad libitum during the experimentperiod The animals were purchased from the Animal Lab-oratory House Wroclaw Medical School and housed in theAnimal Experimental Laboratory (Wroclaw Medical SchoolNorwida 34 Poland) Mice were maintained on a 12 h light-dark cycle at 22 plusmn 02∘C

221 Mice Endurance Exercise on Treadmill The animalsused in this study were divided into two groups sedentarycontrol animals which did not undergo physical activity(119899 = 6) and animals undergoing physical activity (119899 =6) Animals were exercise-trained (119899 = 6) on an Exer 36

Treadmill (Columbus Instruments Columbus OH USA)3 dwk (Monday Wednesday and Friday) for 5wk The micewere accustomed to the treadmill a week before training Forthe 5wk training period mice were subjected to a progressiveexercise protocol with the training portion of the protocolbeginning at 14mmin for 45min (wk 1) and increasingto 24mmin for 45min (wk 5) The training portion ofthe protocol was always preceded by a 10min warm-up at10mmin and followed by a 5min cool-down at 10mmin asdescribed previously [8] SED mice (119899 = 6) were exposedto the treadmill and were given similar inducements on thesame days as EX mice but were not subjected to trainingThe training intensity corresponded to 70ndash75 of VO

2max(murine maximal oxygen uptake) Electrical stimulation wasnot used to encourage the animals to run

222 Mice Exercise on Rotating Wheels Twelve 4-week-oldC57BL6 mice were accustomed for 7 days to the presenceof a rotating wheel and were subsequently subjected tocontrolled 45-minute exercise on rotating wheels Standardmouse exercise wheels were attached directly to the side wallof the cages and each wheel had a magnet directly attachedPrior to commencement of the running trial the mice in theEX group were placed in a cage containing an exercise wheeland kept there for 7 days

223 Human Volunteers Twelve healthy volunteers with amean age of 24 plusmn 1 years (20ndash28 years old) mean bodyweight of 850 plusmn 32 kg (680ndash935 kg) and mean VO

2max of494 plusmn 16mL kgminus1minminus1 (433ndash551mL kgminus1 minminus1) wererecruited to take part in the experiment All individuals weredivided into two groups sedentary (119899 = 6) and physicallyactive students (119899 = 6) The volunteers recruited to thesedentary group were students at Wroclaw Medical Schooland were not engaged in regular training Physically activestudents were recruited from Wroclaw Sport Club (WKSSlask) and had trained in long-distance running for oneyear with endurance training performed six times per weekduring the year The regular training sessions consisted ofendurance running at a distance of 9-10 kmday with a speedof 02 kmmin which is equivalent to sim65 of VO

2maxThe peripheral blood from physically active students wascollected 30min after the last training session in the Faculty ofPharmacy Medical University inWroclaw Following routinemedical screening subjectswere advised of the purpose of thestudy and associated risks and all provided written informedconsent The experimental protocol was approved by theLocal Ethical Committee at Wroclaw Medical School

224 FACS-Based Analysis of VSELs and HSPCs fromMurine BM and PB Total nucleated cells (TNCs) whichwere obtained from BM and PB were subsequentlystained for CD45 hematopoietic lineage markers (Lin)and Sca-1 antigen for 30min in medium containing2 fetal bovine serum The following anti-mouseantibodies (BD Pharmingen) were used for stainingrat anti-CD45 (allophycocyaninCy7 clone 30F11) anti-CD45RB220 (PE clone RA-6B2) anti-Gr-1 (PE cloneRB6-8 C5) anti-T-cell receptor-120572120573 (PE clone H57-5970)

Stem Cells International 3

anti-T-cell receptor-120575 (PE clone GL3) anti-CD11b (PEclone M170) anti-Ter119 (PE clone TER-119) and anti-Ly-6AE (also known as Sca-1 biotin clone E13-1617 withstreptavidin conjugated to PEndashCy5) Cells were then washedresuspended in RPMI medium with 2 fetal bovine serumand sorted with an Influx cell sorter (BD CA USA) Twopopulations were analyzed LinminusSca-1+CD45minus (VSELs) andLinminusSca-1+CD45+ (HSPCs) [18]

225 FACS-Based Analysis of VSELs andHSPCs fromHumanPB Whole human PB was lysed in BD lysing buffer (BDBiosciences USA) for 15min at room temperature andwashed twice in phosphate-buffered saline (PBS) A single-cell suspension was stained for lineage markers (CD2 cloneRPA-210 CD3 clone UCHT1 CD14 clone M5E2 CD66bclone G10F5 CD24 clone ML5 CD56 clone NCAM162CD16 clone 3G8 CD19 clone HIB19 and CD235a clone GA-R2) conjugatedwith fluorescein isothiocyanate (FITC) CD45(cloneHI30) conjugatedwith phycoerythrin (PE) and a com-bination of CD133 (CD1331) conjugated with APC CD34(clone 581) conjugated with PB CD31 (clone WM59) (APC-Cy7) and CD51 (clone 23C6 RUO) conjugated with PE-Cy7for 30min on ice After washing VSELs (CD45minusLinminusCD133+and CD45minusLinminusCD34+ cells) HSPCs (CD45+LinminusCD133+and CD45+LinminusCD34+ cells) EPCs (CD34+CD133+KDR+)and MSCs (CD45minusLinminusCD31minus CD51+) were analyzed byfluorescence-activated cell sorting (FACS Navios BeckmanCoulter USA) At least 106 events were acquired and analyzedusing Kaluza software

226 Real-TimeQuantitative Reverse Transcription PCR (RQ-PCR) Total RNA was isolated from cells harvested from BMandPB from experimental and controlmice by employing theRNeasy Kit (Qiagen Valencia CA) The RNA was reverse-transcribed with MultiScribe reverse transcriptase and oligo-dT primers (Applied Biosystems Foster City CA) Quanti-tative assessment of mRNA levels was performed by real-time reverse transcriptase polymerase chain reaction (RT-PCR) on anABI 7500 Fast instrument employing Power SyBRGreen PCR Master Mix reagent Real-time conditions wereas follows 95∘C (15 sec) followed by 40 cycles at 95∘C (15 sec)and 60∘C (1min) According to melting point analysis onlyone PCR product was amplified under these conditions Therelative quantity of target normalized to the endogenouscontrol 120573-2 microglobulin gene and relative to a calibratoris expressed as fold change (2minusΔΔCt) where ΔCt = (Ct oftarget gene) minus (Ct of the endogenous control gene 120573-2microglobulin) and ΔΔCt = (ΔCt of target gene) minus (ΔCt ofcalibrator for the target gene) The following primer pairswere used 1205732-microglobin 51015840-CAT ACG CCT GCA GAGTTA AGC A-31015840 (forward) and 51015840-GAT CAC ATG TCT CGATCC CAG TAG-31015840 (reverse) Oct-4 51015840-TTC TCA ATG CTAGTT CGC TTT CTC T-31015840 (forward) and 51015840-ACC TTC AGGAGA TAT GCA AAT CG-31015840 (reverse) Sox2 51015840-GCG GAGTGG AAA CTT TTG TCC-31015840 (forward) and 51015840-GGG AAGCGT GTA CTT ATC CTT CT-31015840 (reverse) Rex1 51015840-AGATGG CTT CCC TGA CGG ATA-31015840 (forward) and 51015840-CCTCCA AGC TTT CGA AGG ATT T-31015840 (reverse)

227 In Vitro Clonogenic Assays of Murine HSPCs Thegrowth of murine HSPCs isolated from BM and PB was eval-uated by an in vitro clonogenic assay as described previously[17] Briefly 2 times 105 BM-derived and 4 times 105 PB-derivedcells were resuspended in 04mL of RPMI-1640 mediumand mixed with 18mL of MethoCult HCC-4230 methylcel-lulose medium (StemCell Technologies Inc Canada) sup-plemented with L-glutamine and antibiotics Specific murinerecombinant growth factors (all from RampD Systems USA)were added To stimulate granulocyte-macrophage colony-forming units (CFU-GM) IL-3 (20UmL) SCF (10 ngmL)and GM-CSF (5 ngmL) were used EPO (5UmL) SCF(10 ngmL) and IL-3 (20UmL) were used to stimulateerythrocyte burst-forming units (BFU-E) The colonies werecounted under an inverted microscope after 7ndash10 days of cul-ture Each clonogenic assay was performed in quadruplicate

228 ELISA to Detect Murine MAC SDF-1 and VEGF Fiftymicroliters of PB was taken from the vena cava of the sixC57BL6 mice and collected into Microvette EDTA-coatedtubes (Sarstedt Inc Newton NC)The concentration of C5b-C9 (MAC) was measured by employing the commerciallyavailable highly sensitive enzyme-linked immunosorbentassay (ELISA) kit K-ASSAY (Kamiya Biomedical CompanySeattle WA USA) according to the manufacturerrsquos protocolThe concentrations of SDF-1 and VEGF were measuredby employing the commercially available highly sensitiveenzyme-linked immunosorbent assay (ELISA) (RampD Sys-tems Europe Ltd)

229 ELISA to Detect Human Oct-4 Sox2 and NanogFifty microliters of PB was taken from the vena cava ofthe six C57BL6 mice and collected into Microvette EDTA-coated tubes (Sarstedt Inc Newton NC)The concentrationsof Oct-4 Sox2 and Nanog were measured by employingthe commercially available highly sensitive enzyme-linkedimmunosorbent assay (ELISA) (MyBioSource) accordingto the manufacturerrsquos protocol Results were calculated andpresented as percentage fold difference

2210 Statistical Analysis All data were analyzed usingMicrosoft Excel 2007 and Statistica version 71 software Mostresults are presented as mean plusmn standard error of the meanThe Mann-Whitney 119880 and Studentrsquos 119905-tests were used andstatistical significance was defined as 119875 lt 005

3 Results

31 Short Exercise on Rotating Wheels Mobilizes VSELs intoPeripheral Blood It has been demonstrated that physicalactivity mobilizes HSPCs both in mice and humans [6ndash9]To address the effect of short periods of physical activity onthe mobilization of VSELs we exercised normal mice for45 minutes on a standard rotating wheel and immediatelyafterwardsmicewere evaluated for the numbers ofVSELs andHSPCs circulating in peripheral blood (PB Figures 1(a) and1(b)) and activation of the complement cascade (Figure 1(c))which as we demonstrated previously is crucial for egress ofstem cells from the bone marrow (BM) microenvironment


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Figure 1 Effect of short (45min) exercise on a rotating wheel on stem cell mobilization in mice (a b) The number of VSELs (a) and HSPCs(b) circulating in PB after 45min of exercise on a rotating wheel (effort) compared with nonexercising mice (control) (c) Activation of thecomplement cascade in PB after 45min of exercise on a rotating wheel (effort) compared with nonexercising mice (control) measured byC5b-C9 (MAC) ELISA (d)The increase in the number of clonogenic progenitors circulating in PB after 45min of exercise on rotating wheels(effort) compared with nonexercising mice (control) (119899 = 6micegroup)

into PB [13ndash15] We observed a significant sim2x increase inthe numbers of VSELs (Figure 1(a)) increased numbers ofHSPCs (Figure 1(b)) and increased numbers of clonogenicCFU-GM circulating in PB (Figure 1(d)) The increases innumber of these circulating cells correlated with activationof the complement cascade as evidenced by an increase inC5bC9 (also known as themembrane attack complex (MAC))measured in PB plasma by ELISA (Figure 1(c))

32 Prolonged Endurance Exercise on a Treadmill Increasesthe Number of VSELs Circulating in PB as well as Residingin BM Subsequently we performed an endurance exerciseexperiment in which mice were subjected to forced running

on a treadmill for 5 days or 5 weeks Figure 2(a) shows thatforced exercise for 5 consecutive days or 5 weeks significantlyenhanced the number of VSELs both circulating in PB (leftpanel) and residing in BM (right panel) This increase inthe number of VSELs circulating in PB correlated with anincrease in expression of mRNA for VSEL markers suchas Oct-4 Sox2 and Nanog [11 12] in PB mononuclearcells (Figure 2(b)) Again mobilization of VSELs correlatedwith an increase in activation of the complement cascade(Figure 2(c)) A representative FACS analysis of VSELs circu-lating in PB in control mice as well as mice that had exercisedfor 5 days or 5weeks is shown in Supplemental Digital Contentavailable online at httpdxdoiorg1011552015395971


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Figure 2 Effect of 5 days or 5 weeks of exercise on a treadmill on the number of VSELs in PB and BM (a)The number of VSELs circulating inPB (left panel) and BM (right panel) after 5 days or 5 weeks of exercise on a treadmill (effort) compared with nonexercisingmice (control) (b)Quantitative real-time PCR changes in expression of Oct4 Sox2 and Nanog in PB mononuclear cells after 5 days or 5 weeks of exercise on atreadmill (effort) compared with nonexercising mice (control) whose expression level was defined as 100 (c) Activation of the complementcascade in PB measured by C5b-C9 (MAC) ELISA after 5-week exercise on a treadmill (effort) compared with nonexercising mice (control)(119899 = 6micegroup)

Interestingly these changes in the numbers of VSELs andHSPCs did not correlate with the plasma levels of SDF-1or VEGF In fact we did not observe significant changesin the levels of these factors in PB between control and

exercising mice (data not shown) which indicates alongwith our previous observations in other models of stem cellmobilization that if the complement cascade is activatedand stem cells are released from their niches in BM due to


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5-day effort 5-week effort 5-day effort 5-week effort

(b)

Figure 3 Effect of 5 days or 5 weeks of exercise on a treadmill on the number of HSPCs in PB and BM (a)The number of HSPCs circulatingin PB (left panel) and BM (right panel) after 5 days or 5 weeks of exercise on a treadmill (effort) compared with nonexercising mice (control)(b) The increase in the number of clonogenic progenitors (CFU-GM left panel and BFU-E right panel) in BM after 5 days or 5 weeks ofexercise on a treadmill (effort) compared with nonexercising mice (control) (119899 = 6micegroup)

a proteolytic microenvironment the plasma level of S1P issufficiently high to induce egress of stem cells from BM intoPB [14]

At the same time we observed the predicted increase inHSPCs (Figure 3(a)) both circulating in PB (left panel) andresiding in BM (right panel) An increase in the numbers ofHSPCs in BM correlated with increases in the numbers ofclonogenic CFU-GM and BFU-E progenitors (Figure 3(b))

33 Increases in VSELs Circulating in PB after RunningExercise in Young Athletes The effect of running exercise

on increasing the numbers of HSPCs circulating in PBhas already been demonstrated in humans [8] In thisstudy encouraged by our murine data we enumerated thenumber of small CD34+LinminusCD45minus and CD133+LinminusCD45minuscells circulating in PB (Figure 4(a)) that correspond to theVSEL population (left panels) in sedentary and physicallyactive students and in parallel we enumerated the num-ber of CD34+LinminusCD45+ and CD133+LinminusCD45+ cells thatcorrespond to HSPCs (right panels) Figure 4(a) shows anincrease in the numbers of these cells circulating in PB inphysically active subjects after running effort (90ndash120min of


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Figure 4 Effect of exercise on stem cell mobilization in healthy young volunteers (a) The number of CD133+ and CD34+ VSELs (left)and CD133+ and CD34+ HSPCs (right) circulating in PB in young athletes after not exercising (control) or after running exercise (effort) (b)Quantitative real-time PCR changes in expression of Oct4 Sox2 andNanog in PBmononuclear cells after running exercise (effort) comparedwith nonexercising subjects (control) Changes are shown compared with values detected in control volunteers (defined as 100) (c) Thenumber of EPCs (left panel) andMSCs (right panel) circulating in PB in young athletes after not exercising (control) or after running exercise(effort) (119899 = 6 volunteersgroup)


training session corresponding to 65 of training consistingof VO

2peak at the distance of 8 km) An increase in the numberof VSELs circulating in PB has been subsequently con-firmed by ELISA to detect Oct-4 Sox2 and Nanog proteins(Figure 4(b)) We also observed a small increase in the num-ber of EPCs but not MSCs circulating in PB (Figure 4(c))

4 Discussion

The salient observation of this work is the positive effectof exercise on mobilization into PB and expansion in BMof VSELs Since these small developmentally early cellsmay differentiate into several types of cells across all threegerm layers and play a role in regeneration [10ndash12 18] thisobservation may explain in a novel way the positive effect ofregular exercise on tissue and organ rejuvenation

In addition to VSELs we have confirmed that physicalactivity also mobilizes HSPCs into PB In support of thisobservation there are several reports in the literature demon-strating that physical exercise mobilizes both HSPCs andEPCs into PB [6 8 9] The efficacy of the mobilization mayvary depending on the intensity of the exercise protocolsemployed in humanor animal subjects and the time of samplecollection of PB or BM after exercise for enumeration ofthe stem cells In our experiments we evaluated the effectof exercise on the release of VSELs and HSPCs from BMimmediately after a short intensive run on rotating wheels orafter repeated daily running exercise on a treadmill for 5 daysor 5 weeks We also evaluated the mobilization of VSELs andHSPCs in healthy young athletes after a 10 km run It is mostlikely that stem cells mobilized during strenuous exercise actas ldquocirculating paramedicsrdquo with a role in repairing micro-scopic damage in skeletal muscles as well as in other tissuesAn important role in this phenomenon may be played byVSELs which in appropriate in vivo models are reportedlyable to differentiate into cells for different tissues includingmesenchymal precursors lung alveolar epithelium gametesand endothelial cells [19ndash22] In vitro research proved thatnot only does an enabling environment improve releasingand increasing cellsrsquo proliferation but both mechanical andphysical stress as well changed conditions (ie differentculture surfaces) are associated withmechanisms controllingcellular functions These findings provide evidence thatendurance training may play modulating role in regulationof cellular functions and VSELs releasing [23ndash25]This abilitycorresponds with several observations in which VSELs arereleased into PB in clinical situations of organ or tissuedamage such as heart infarct [26] stroke [27] skin burns[28] and neural tissue toxic damage [29] and the extent oftheir mobilization into PB may even have some prognosticvalue [26ndash28] Our data presented herein show for the firsttime that not only are VSELs released from BM into PB inresponse to physical exercise but there is also a positive effectof exercise on the expansion of this primitive pool of stemcells in BM

We also observed that endurance exercise increases thenumber of HSPCs in BM corroborating reports from otherinvestigators [7 9] Specifically mice trained on a treadmill at

progressive speeds over a 10-week period displayed increasedmedullary and mobilized HSPC content from 50 to 800depending on the HSPC type and marrow cavity fat wasreduced by 78 [6] Interestingly as has been demonstratedin another paper from this group exercise promoted BM cellsurvival and exercise training increased survival of recipientmice after BM transplantation with increased total bloodcell reconstitution [30] Based on this finding endurancetraining has a positive prohematopoietic effect both directlyon HSPCs and on accessory cells in the BM microenvi-ronment (eg stroma and osteoblasts) that provide nichesfor these cells [4] Interestingly in addition to HSPCs ithas been reported that physical activity also has a positiveeffect on neurogenesis in the adult and aging brain [31 32]Thus physical activity has a positive effect on several typesof tissue-committed stem cells in various organs such asskeletal muscle satellite stem cells [31 33] HSPCs [9] andneural progenitors and several factors triggered by exercisesuch as insulin-like growth factor 1 VEGF platelet-derivedgrowth factor hepatocyte growth factor and hormones suchas androgens mediate this effect [31] On the other handit is very likely that other non-peptide-based regulators ofcell growth such as bioactive phosphosphingolipids andalarmines released from hypoxic tissues may play an impor-tant role as well [31]

Mobilization into PB and expansion in BM of VSELswhich may differentiate into several types of cells across thethree germ layers [10 11 19ndash22] and express several genescharacteristic of early development stem cells (Oct-4 Sox-2 and Nanog) may explain in a novel way the positiveeffect of regular exercise on tissue and organ rejuvenationOf note there is a similar positive effect on this pool of stemcells by caloric restriction as we demonstrated recently [16]Therefore VSELs may reconcile all observations reported sofar of a positive effect of both exercise and caloric restrictionon life quality and the extension of life span [12] However weare aware that more direct data are necessary to fully supportthis novel concept

Mobilization of both HSPCs and VSELs correlated in ourstudies with activation of the complement cascadeThis againconfirms the pivotal role of this cascade which is also seenas a response to infection or tissue and organ damage intriggering the mobilization process after strenuous exercise[13] Since the complement cascade has robust cross-talk withtwo other evolutionarily ancient proteolytic cascadesmdashthecoagulation and fibrinolytic cascades [34]mdashfurther studiesare needed to assess the role of the products of activation ofthese cascades on mobilization of stem cells in response toendurance exercise

We conclude that physical activity may have a positiveeffect on improving life quality by directly affecting thepool of the most primitive stem cells residing in adulttissues Future studies will be important to address thequestion of whether a combination of physical activity withcaloric restriction and some pharmacological drugs suchas metformin which is currently employed to increase lifespan have a synergistic effect on VSELs and tissue andorgan rejuvenationThese observations will be crucial for thedevelopment and optimization of novel treatment strategies


aimed at prolonging human life span and physical activity isan important part of this

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

This work was supported by NIHGrants 2R01 DK074720 andR01HL112788 the Stella andHenry Endowment andMaestroGrant 201102ANZ400035 to Mariusz Z Ratajczak Thiswork was supported by Wroclaw Research Centre EIT+under Project ldquoBiotechnologies and AdvancedMedical Tech-nologiesrdquo BioMed (POIG010102-02-00308) financed fromthe EuropeanRegional Development Fund (Operational Pro-grammed 10 Innovative Economy 112) to KrzysztofMaryczThe authors acknowledge the Leading National ResearchCentre ldquoKNOWrdquo (Krajowy Naukowy Osrodek Wiodący) forcovering the publication fee

References

[1] H Bonig and T Papayannopoulou ldquoHematopoietic stem cellmobilization updated conceptual renditionsrdquoLeukemia vol 27no 1 pp 24ndash31 2013

[2] M Z Ratajczak T Jadczyk D Pędziwiatr andWWojakowskildquoNew advances in stem cell research practical implica-tions for regenerative medicinerdquo Polskie Archiwum MedycynyWewnętrznej vol 124 pp 417ndash426 2014

[3] M Z Ratajczak K Marycz A Poniewierska-Baran KFiedorowicz M Zbucka-Kretowska and M Moniuszko ldquoVerysmall embryonic-like stem cells as a novel developmental con-cept and the hierarchy of the stem cell compartmentrdquo Advancesin Medical Sciences vol 59 no 2 pp 273ndash280 2014

[4] T Lapidot and O Kollet ldquoThe brain-bone-blood triad trafficlights for stem-cell homing and mobilizationrdquo HematologytheEducation Program of the American Society of Hematology vol2010 pp 1ndash6 2010

[5] SMassberg P Schaerli I Knezevic-Maramica et al ldquoImmuno-surveillance by hematopoietic progenitor cells traffickingthrough blood lymph and peripheral tissuesrdquo Cell vol 131 no5 pp 994ndash1008 2007

[6] J M Baker M de Lisio and G Parise ldquoEndurance exercisetraining promotes medullary hematopoiesisrdquo The FASEB Jour-nal vol 25 no 12 pp 4348ndash4357 2011

[7] M de Lisio and G Parise ldquoCharacterization of the effects ofexercise training on hematopoietic stem cell quantity andfunctionrdquo Journal of Applied Physiology vol 113 no 10 pp 1576ndash1584 2012

[8] J M Kroepfl K Pekovits I Stelzer et al ldquoExercise increasesthe frequency of circulating hematopoietic progenitor cells butreduces hematopoietic colony-forming capacityrdquo StemCells andDevelopment vol 21 no 16 pp 2915ndash2925 2012

[9] K A Volaklis S P Tokmakidis and M Halle ldquoAcute andchronic effects of exercise on circulating endothelial progenitorcells in healthy and diseased patientsrdquo Clinical Research inCardiology vol 102 no 4 pp 249ndash257 2013

[10] A M Havens H Sun Y Shiozawa et al ldquoHuman and murinevery small embryonic-like cells represent multipotent tissueprogenitors in vitro and in vivordquo Stem Cells and Developmentvol 23 no 7 pp 689ndash701 2014

[11] M Kucia M Wysoczynski J Ratajczak and M Z RatajczakldquoIdentification of very small embryonic like (VSEL) stem cells inbone marrowrdquo Cell and Tissue Research vol 331 no 1 pp 125ndash134 2008

[12] M Z Ratajczak D-M Shin R Liu et al ldquoVery SmallEmbryonicEpiblast-Like stem cells (VSELs) and their potentialrole in aging and organ rejuvenationmdashan update and compar-ison to other primitive small stem cells isolated from adulttissuesrdquo Aging vol 4 no 4 pp 235ndash246 2012

[13] H M Lee W Wu M Wysoczynski et al ldquoImpaired mobi-lization of hematopoietic stemprogenitor cells in C5-deficientmice supports the pivotal involvement of innate immunity inthis process and reveals novel promobilization effects of granu-locytesrdquo Leukemia vol 23 no 11 pp 2052ndash2062 2009

[14] M Z Ratajczak H Lee M Wysoczynski et al ldquoNovel insightinto stem cell mobilization-plasma sphingosine-1-phosphate isamajor chemoattractant that directs the egress of hematopoieticstem progenitor cells from the bone marrow and its level inperipheral blood increases during mobilization due to acti-vation of complement cascademembrane attack complexrdquoLeukemia vol 24 no 5 pp 976ndash985 2010

[15] S Rafii B Heissig and K Hattori ldquoEfficient mobilization andrecruitment of marrow-derived endothelial and hematopoieticstem cells by adenoviral vectors expressing angiogenic factorsrdquoGene Therapy vol 9 no 10 pp 631ndash641 2002

[16] MWysoczynski J RatajczakD Pedziwiatr G Rokosh R Bolliand M Z Ratajczak ldquoIdentification of heme oxygenase 1 (HO-1) as a novel negative regulator ofmobilization of hematopoieticstemprogenitor cellsrdquo StemCell Reviews andReports vol 11 no1 pp 110ndash118 2015

[17] K Grymula K Piotrowska S Słuczanowska-Głąbowska et alldquoPositive effects of prolonged caloric restriction on the popu-lation of very small embryonic-like stem cellsmdashhematopoieticand ovarian implicationsrdquo Journal of Ovarian Research vol 7no 1 article 68 2014

[18] M Suszynska E K Zuba-Surma M Maj et al ldquoThe propercriteria for identification and sorting of very small embryonic-like stem cells and some nomenclature issuesrdquo Stem Cells andDevelopment vol 23 no 7 pp 702ndash713 2014

[19] S H Kassmer H Jin P-X Zhang et al ldquoVery small embryonic-like stem cells from the murine bone marrow differentiate intoepithelial cells of the lungrdquo Stem Cells vol 31 no 12 pp 2759ndash2766 2013

[20] S H Kassmer E M Bruscia P X Zhang and D S KrauseldquoNonhematopoietic cells are the primary source of bonemarrow-derived lung epithelial cellsrdquo Stem Cells vol 30 no 3pp 491ndash499 2012

[21] I Virant-Klun N Zech P Rozman et al ldquoPutative stem cellswith an embryonic character isolated from the ovarian surfaceepithelium of women with no naturally present follicles andoocytesrdquo Differentiation vol 76 no 8 pp 843ndash856 2008

[22] D Bhartiya S Kasiviswananthan and A Shaikh ldquoCellularorigin of testis-derived pluripotent stem cells a case for verysmall embryonic-like stem cellsrdquo Stem Cells and Developmentvol 21 no 5 pp 670ndash674 2012

[23] M Maredziak K Marycz A Smieszek D Lewandowski andN Y Toker ldquoThe influence of static magnetic fields on canineand equine mesenchymal stem cells derived from adipose


tissuerdquo In Vitro Cellular amp Developmental Biology Animal vol50 no 6 pp 562ndash571 2014

[24] A Donesz-Sikorska K Marycz A Smieszek J Grzesiak KKalinski and J Krzak-Ros ldquoBiologically active oxide coatingsproduced by the sol-gel method on steel implantrdquo PrzemysłChemiczny vol 92 no 6 pp 1110ndash1116 2013

[25] K Marycz J Krzak-Ros A Smieszek J Grzesiak and ADonesz-Sikorska ldquoEffect of oxide materials synthesized withsolndashgel method on adhesion of mesenchymal stem cellsrdquoPrzemysl Chemiczny vol 92 no 6 pp 1000ndash1003 2013

[26] W Wojakowski M Tendera M Kucia et al ldquoMobilization ofbone marrow-derived Oct-4+ SSEA-4+ very small embryonic-like stem cells in patients with acute myocardial infarctionrdquoJournal of the American College of Cardiology vol 53 no 1 pp1ndash9 2009

[27] E Paczkowska M Kucia D Koziarska et al ldquoClinical evidencethat very small embryonic-like stem cells are mobilized intoperipheral blood in patients after strokerdquo Stroke vol 40 no 4pp 1237ndash1244 2009

[28] J Drukała E PaczkowskaM Kucia et al ldquoStem cells includinga population of very small embryonic-like stem cells aremobilized into peripheral blood in patients after skin burninjuryrdquo Stem Cell Reviews and Reports vol 8 no 1 pp 184ndash1942012

[29] K Grymula M Tarnowski K Piotrowska et al ldquoEvidence thatthe population of quiescent bone marrow-residing very smallembryonicepiblast-like stem cells (VSELs) expands in responseto neurotoxic treatmentrdquo Journal of Cellular and MolecularMedicine vol 18 no 9 pp 1797ndash1806 2014

[30] M de Lisio J M Baker and G Parise ldquoExercise promotes bonemarrow cell survival and recipient reconstitution post-bonemarrow transplantation which is associated with increasedsurvivalrdquo Experimental Hematology vol 41 no 2 pp 143ndash1542013

[31] F MacAluso and K H Myburgh ldquoCurrent evidence thatexercise can increase the number of adult stem cellsrdquo Journal ofMuscle Research and Cell Motility vol 33 no 3-4 pp 187ndash1982012

[32] K Fabel S A Wolf D Ehninger H Babu P Leal-Galiciaand G Kempermann ldquoAdditive effects of physical exercise andenvironmental enrichment on adult hippocampal neurogenesisin micerdquo Frontiers in Neuroscience vol 3 article 50 2009

[33] T Snijders L B Verdijk J S Smeets et al ldquoThe skeletal musclesatellite cell response to a single bout of resistance-type exerciseis delayed with aging in menrdquo Age vol 36 no 4 p 9699 2014

[34] S Borkowska M Suszynska K Mierzejewska et al ldquoNovelevidence that crosstalk between the complement coagulationand fibrinolysis proteolytic cascades is involved in mobilizationof hematopoietic stemprogenitor cells (HSPCs)rdquo Leukemiavol 28 pp 2148ndash2154 2014

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


Egress of stem cells from the BM is triggered by activa-tion of the complement cascade which releases importantactive cleavage fragments such as the C5 component C5athat induce granulocytes and monocytes in BM to releaseproteolytic enzymes that attenuate stem cell retention signalsin BM niches and permeabilize the BMndashPB barrier thusfacilitating the egress of stem cells [13] The major chemoat-tractant for stem cells in PB is sphingosine-1-phosphate (S1P)and in the process of mobilization certain other factors arealso involved including 120572-chemokine stromal-derived factor1 (SDF-1) [14] It has also been proposed that mobilizationof particular types of stem cells may be modulated by stemcell type-specific factors such as vascular endothelial growthfactor (VEGF) which is involved in mobilization of EPCs[15] Our recent research demonstrated that mobilization ofstem cells not only is triggered by but also correlates withactivation of the complement cascade and its activation canbe easilymeasured in PB using commercially available ELISAassays to detect for example the C5b-C9 membrane attackcomplex (MAC) [16]

Our data confirm that HSPCs are released from BMinto PB in response to physical exercise and demonstratefor the first time that in parallel VSELs are also mobilizedImportantly we demonstrate a positive effect of exercise onexpansion of this primitive pool of stem cells in BM Sincethese small cells may differentiate into several types of cellsacross the three germ layers their increase may explain ina novel way the positive effect of regular exercise on tissueand organ rejuvenation Of note a similar positive effecton this pool of cells by caloric restriction has recently beendemonstrated [17]Therefore VSELsmay be able to reconcilereported observations of a positive effect of both exercise andcaloric restriction on life quality and extension of life spanHowever we are aware that more direct data are necessary tofully support this novel concept

2 Materials and Methods

21 Animal Care and Use Conducted studies were approvedby the II Local Ethical Committee Environmental and LifeScience University and adhered to American College ofSports Medicine (ACSM) animal care standards

22 Experimental Animals Theexperiments were performedon 90 4-week-old C57BL6 mice Sedentary control (SED)and exercise-trained (EX) C57Bl6 mice were housed threeper cage in an ultraclean facility on ventilated racks and wereprovided food and water ad libitum during the experimentperiod The animals were purchased from the Animal Lab-oratory House Wroclaw Medical School and housed in theAnimal Experimental Laboratory (Wroclaw Medical SchoolNorwida 34 Poland) Mice were maintained on a 12 h light-dark cycle at 22 plusmn 02∘C

221 Mice Endurance Exercise on Treadmill The animalsused in this study were divided into two groups sedentarycontrol animals which did not undergo physical activity(119899 = 6) and animals undergoing physical activity (119899 =6) Animals were exercise-trained (119899 = 6) on an Exer 36

Treadmill (Columbus Instruments Columbus OH USA)3 dwk (Monday Wednesday and Friday) for 5wk The micewere accustomed to the treadmill a week before training Forthe 5wk training period mice were subjected to a progressiveexercise protocol with the training portion of the protocolbeginning at 14mmin for 45min (wk 1) and increasingto 24mmin for 45min (wk 5) The training portion ofthe protocol was always preceded by a 10min warm-up at10mmin and followed by a 5min cool-down at 10mmin asdescribed previously [8] SED mice (119899 = 6) were exposedto the treadmill and were given similar inducements on thesame days as EX mice but were not subjected to trainingThe training intensity corresponded to 70ndash75 of VO

2max(murine maximal oxygen uptake) Electrical stimulation wasnot used to encourage the animals to run

222 Mice Exercise on Rotating Wheels Twelve 4-week-oldC57BL6 mice were accustomed for 7 days to the presenceof a rotating wheel and were subsequently subjected tocontrolled 45-minute exercise on rotating wheels Standardmouse exercise wheels were attached directly to the side wallof the cages and each wheel had a magnet directly attachedPrior to commencement of the running trial the mice in theEX group were placed in a cage containing an exercise wheeland kept there for 7 days

223 Human Volunteers Twelve healthy volunteers with amean age of 24 plusmn 1 years (20ndash28 years old) mean bodyweight of 850 plusmn 32 kg (680ndash935 kg) and mean VO

2max of494 plusmn 16mL kgminus1minminus1 (433ndash551mL kgminus1 minminus1) wererecruited to take part in the experiment All individuals weredivided into two groups sedentary (119899 = 6) and physicallyactive students (119899 = 6) The volunteers recruited to thesedentary group were students at Wroclaw Medical Schooland were not engaged in regular training Physically activestudents were recruited from Wroclaw Sport Club (WKSSlask) and had trained in long-distance running for oneyear with endurance training performed six times per weekduring the year The regular training sessions consisted ofendurance running at a distance of 9-10 kmday with a speedof 02 kmmin which is equivalent to sim65 of VO

2maxThe peripheral blood from physically active students wascollected 30min after the last training session in the Faculty ofPharmacy Medical University inWroclaw Following routinemedical screening subjectswere advised of the purpose of thestudy and associated risks and all provided written informedconsent The experimental protocol was approved by theLocal Ethical Committee at Wroclaw Medical School

224 FACS-Based Analysis of VSELs and HSPCs fromMurine BM and PB Total nucleated cells (TNCs) whichwere obtained from BM and PB were subsequentlystained for CD45 hematopoietic lineage markers (Lin)and Sca-1 antigen for 30min in medium containing2 fetal bovine serum The following anti-mouseantibodies (BD Pharmingen) were used for stainingrat anti-CD45 (allophycocyaninCy7 clone 30F11) anti-CD45RB220 (PE clone RA-6B2) anti-Gr-1 (PE cloneRB6-8 C5) anti-T-cell receptor-120572120573 (PE clone H57-5970)









3 Results



VSELs

0

200

400

600lowastP lt 005

Num

ber o

f cel

ls1

mL

perip

hera

l blo

od

Control Effort

(a)

0

2501250

1500

1750

2000

HSPCslowastP lt 005

Control Effort

Num

ber o

f cel

ls1

mL

perip

hera

l blo

od

(b)

MAC

00

05

10

15lowastP lt 005

MAC

(ng

mL)

Control Effort

(c)

0

20

40

60

80

100

120

140

160


Num

ber o

f CFU

-GM

105

cells

Control Effort

(d)






VSELs

0

200

400

600

800

1000

VSELs

0

10

20

30

40

50



Num

ber o

fLin

minusSc

a-1+

CD45

minus1

mL

perip

hera

l blo

od


Num

ber o

fLin

minusSc

a-1+

CD45

minus

106

MN

Cs

(a)

Oct4

0

50

100

150

200

250

300

350

Fold

diff

eren

ce (

)

5 days 5 weeks

Sox2

0

40

80

120360

400

440

Fold

diff

eren

ce (

)

Nanog

0

4080

120

160

Fold

diff

eren

ce (





(b)

MAC

00

05

10

15

5 weeks

MAC

(ng

mL)

Control Effort

lowastP lt 005

(c)





HSPCs

0

1000

2000

3000HSPC MNC

0

50

100

150

200




Num

ber H

SPCs

1m

L pe

riphe

ral b

lood

Num

ber H

SPCs

1m

in M

NCs

(a)

CFU-GM

0

100

200

300

Fold

diff

eren

ce (

)

0

100

200

300

400

500

BFU-EFo

ld d

iffer

ence

()

Control Control



(b)







000

002

004

006

008

00

05

10

15

000

002

004

006

008

010

012

00

05

10

15

20

Num

ber o

f CD133+

VSEL

s1120583

Lpe

riphe

ral b

lood

perip

hera

l blo

od

perip

hera

l blo

odpe

riphe

ral b

lood

Num

ber o

f CD133+

HSP

Cs1120583

L

Num

ber o

f CD34+

VSEL

s1120583

L

Num

ber o

f CD34+

HSP

Cs1120583

L

lowastP lt 005

lowastP lt 005

lowastP lt 005

lowastP lt 005





(a)

0

100

200

300

400

500

Fold

diff

eren

ce (

)

Oct4 Sox2

0

100

200

300

400

Fold

diff

eren

ce (

)

Nanog

0

50

100

150

Fold

diff

eren

ce (

)


lowastP lt 005


(b)

EPCs

00

01

02

03

04

05MSCs

00

02

04

06

08

perip

hera

l blo

od

perip

hera

l blo

od



Num

ber o

f EPC

s1120583

L

Num

ber o

f MSC

s1120583

L

(c)





4 Discussion












Acknowledgments


References












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology









3 Results



VSELs

0

200

400

600lowastP lt 005

Num

ber o

f cel

ls1

mL

perip

hera

l blo

od

Control Effort

(a)

0

2501250

1500

1750

2000

HSPCslowastP lt 005

Control Effort

Num

ber o

f cel

ls1

mL

perip

hera

l blo

od

(b)

MAC

00

05

10

15lowastP lt 005

MAC

(ng

mL)

Control Effort

(c)

0

20

40

60

80

100

120

140

160


Num

ber o

f CFU

-GM

105

cells

Control Effort

(d)






VSELs

0

200

400

600

800

1000

VSELs

0

10

20

30

40

50



Num

ber o

fLin

minusSc

a-1+

CD45

minus1

mL

perip

hera

l blo

od


Num

ber o

fLin

minusSc

a-1+

CD45

minus

106

MN

Cs

(a)

Oct4

0

50

100

150

200

250

300

350

Fold

diff

eren

ce (

)

5 days 5 weeks

Sox2

0

40

80

120360

400

440

Fold

diff

eren

ce (

)

Nanog

0

4080

120

160

Fold

diff

eren

ce (





(b)

MAC

00

05

10

15

5 weeks

MAC

(ng

mL)

Control Effort

lowastP lt 005

(c)





HSPCs

0

1000

2000

3000HSPC MNC

0

50

100

150

200




Num

ber H

SPCs

1m

L pe

riphe

ral b

lood

Num

ber H

SPCs

1m

in M

NCs

(a)

CFU-GM

0

100

200

300

Fold

diff

eren

ce (

)

0

100

200

300

400

500

BFU-EFo

ld d

iffer

ence

()

Control Control



(b)







000

002

004

006

008

00

05

10

15

000

002

004

006

008

010

012

00

05

10

15

20

Num

ber o

f CD133+

VSEL

s1120583

Lpe

riphe

ral b

lood

perip

hera

l blo

od

perip

hera

l blo

odpe

riphe

ral b

lood

Num

ber o

f CD133+

HSP

Cs1120583

L

Num

ber o

f CD34+

VSEL

s1120583

L

Num

ber o

f CD34+

HSP

Cs1120583

L

lowastP lt 005

lowastP lt 005

lowastP lt 005

lowastP lt 005





(a)

0

100

200

300

400

500

Fold

diff

eren

ce (

)

Oct4 Sox2

0

100

200

300

400

Fold

diff

eren

ce (

)

Nanog

0

50

100

150

Fold

diff

eren

ce (

)


lowastP lt 005


(b)

EPCs

00

01

02

03

04

05MSCs

00

02

04

06

08

perip

hera

l blo

od

perip

hera

l blo

od



Num

ber o

f EPC

s1120583

L

Num

ber o

f MSC

s1120583

L

(c)





4 Discussion












Acknowledgments


References












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


VSELs

0

200

400

600lowastP lt 005

Num

ber o

f cel

ls1

mL

perip

hera

l blo

od

Control Effort

(a)

0

2501250

1500

1750

2000

HSPCslowastP lt 005

Control Effort

Num

ber o

f cel

ls1

mL

perip

hera

l blo

od

(b)

MAC

00

05

10

15lowastP lt 005

MAC

(ng

mL)

Control Effort

(c)

0

20

40

60

80

100

120

140

160


Num

ber o

f CFU

-GM

105

cells

Control Effort

(d)






VSELs

0

200

400

600

800

1000

VSELs

0

10

20

30

40

50



Num

ber o

fLin

minusSc

a-1+

CD45

minus1

mL

perip

hera

l blo

od


Num

ber o

fLin

minusSc

a-1+

CD45

minus

106

MN

Cs

(a)

Oct4

0

50

100

150

200

250

300

350

Fold

diff

eren

ce (

)

5 days 5 weeks

Sox2

0

40

80

120360

400

440

Fold

diff

eren

ce (

)

Nanog

0

4080

120

160

Fold

diff

eren

ce (





(b)

MAC

00

05

10

15

5 weeks

MAC

(ng

mL)

Control Effort

lowastP lt 005

(c)





HSPCs

0

1000

2000

3000HSPC MNC

0

50

100

150

200




Num

ber H

SPCs

1m

L pe

riphe

ral b

lood

Num

ber H

SPCs

1m

in M

NCs

(a)

CFU-GM

0

100

200

300

Fold

diff

eren

ce (

)

0

100

200

300

400

500

BFU-EFo

ld d

iffer

ence

()

Control Control



(b)







000

002

004

006

008

00

05

10

15

000

002

004

006

008

010

012

00

05

10

15

20

Num

ber o

f CD133+

VSEL

s1120583

Lpe

riphe

ral b

lood

perip

hera

l blo

od

perip

hera

l blo

odpe

riphe

ral b

lood

Num

ber o

f CD133+

HSP

Cs1120583

L

Num

ber o

f CD34+

VSEL

s1120583

L

Num

ber o

f CD34+

HSP

Cs1120583

L

lowastP lt 005

lowastP lt 005

lowastP lt 005

lowastP lt 005





(a)

0

100

200

300

400

500

Fold

diff

eren

ce (

)

Oct4 Sox2

0

100

200

300

400

Fold

diff

eren

ce (

)

Nanog

0

50

100

150

Fold

diff

eren

ce (

)


lowastP lt 005


(b)

EPCs

00

01

02

03

04

05MSCs

00

02

04

06

08

perip

hera

l blo

od

perip

hera

l blo

od



Num

ber o

f EPC

s1120583

L

Num

ber o

f MSC

s1120583

L

(c)





4 Discussion












Acknowledgments


References












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


VSELs

0

200

400

600

800

1000

VSELs

0

10

20

30

40

50



Num

ber o

fLin

minusSc

a-1+

CD45

minus1

mL

perip

hera

l blo

od


Num

ber o

fLin

minusSc

a-1+

CD45

minus

106

MN

Cs

(a)

Oct4

0

50

100

150

200

250

300

350

Fold

diff

eren

ce (

)

5 days 5 weeks

Sox2

0

40

80

120360

400

440

Fold

diff

eren

ce (

)

Nanog

0

4080

120

160

Fold

diff

eren

ce (





(b)

MAC

00

05

10

15

5 weeks

MAC

(ng

mL)

Control Effort

lowastP lt 005

(c)





HSPCs

0

1000

2000

3000HSPC MNC

0

50

100

150

200




Num

ber H

SPCs

1m

L pe

riphe

ral b

lood

Num

ber H

SPCs

1m

in M

NCs

(a)

CFU-GM

0

100

200

300

Fold

diff

eren

ce (

)

0

100

200

300

400

500

BFU-EFo

ld d

iffer

ence

()

Control Control



(b)







000

002

004

006

008

00

05

10

15

000

002

004

006

008

010

012

00

05

10

15

20

Num

ber o

f CD133+

VSEL

s1120583

Lpe

riphe

ral b

lood

perip

hera

l blo

od

perip

hera

l blo

odpe

riphe

ral b

lood

Num

ber o

f CD133+

HSP

Cs1120583

L

Num

ber o

f CD34+

VSEL

s1120583

L

Num

ber o

f CD34+

HSP

Cs1120583

L

lowastP lt 005

lowastP lt 005

lowastP lt 005

lowastP lt 005





(a)

0

100

200

300

400

500

Fold

diff

eren

ce (

)

Oct4 Sox2

0

100

200

300

400

Fold

diff

eren

ce (

)

Nanog

0

50

100

150

Fold

diff

eren

ce (

)


lowastP lt 005


(b)

EPCs

00

01

02

03

04

05MSCs

00

02

04

06

08

perip

hera

l blo

od

perip

hera

l blo

od



Num

ber o

f EPC

s1120583

L

Num

ber o

f MSC

s1120583

L

(c)





4 Discussion












Acknowledgments


References












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


HSPCs

0

1000

2000

3000HSPC MNC

0

50

100

150

200




Num

ber H

SPCs

1m

L pe

riphe

ral b

lood

Num

ber H

SPCs

1m

in M

NCs

(a)

CFU-GM

0

100

200

300

Fold

diff

eren

ce (

)

0

100

200

300

400

500

BFU-EFo

ld d

iffer

ence

()

Control Control



(b)







000

002

004

006

008

00

05

10

15

000

002

004

006

008

010

012

00

05

10

15

20

Num

ber o

f CD133+

VSEL

s1120583

Lpe

riphe

ral b

lood

perip

hera

l blo

od

perip

hera

l blo

odpe

riphe

ral b

lood

Num

ber o

f CD133+

HSP

Cs1120583

L

Num

ber o

f CD34+

VSEL

s1120583

L

Num

ber o

f CD34+

HSP

Cs1120583

L

lowastP lt 005

lowastP lt 005

lowastP lt 005

lowastP lt 005





(a)

0

100

200

300

400

500

Fold

diff

eren

ce (

)

Oct4 Sox2

0

100

200

300

400

Fold

diff

eren

ce (

)

Nanog

0

50

100

150

Fold

diff

eren

ce (

)


lowastP lt 005


(b)

EPCs

00

01

02

03

04

05MSCs

00

02

04

06

08

perip

hera

l blo

od

perip

hera

l blo

od



Num

ber o

f EPC

s1120583

L

Num

ber o

f MSC

s1120583

L

(c)





4 Discussion












Acknowledgments


References












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


000

002

004

006

008

00

05

10

15

000

002

004

006

008

010

012

00

05

10

15

20

Num

ber o

f CD133+

VSEL

s1120583

Lpe

riphe

ral b

lood

perip

hera

l blo

od

perip

hera

l blo

odpe

riphe

ral b

lood

Num

ber o

f CD133+

HSP

Cs1120583

L

Num

ber o

f CD34+

VSEL

s1120583

L

Num

ber o

f CD34+

HSP

Cs1120583

L

lowastP lt 005

lowastP lt 005

lowastP lt 005

lowastP lt 005





(a)

0

100

200

300

400

500

Fold

diff

eren

ce (

)

Oct4 Sox2

0

100

200

300

400

Fold

diff

eren

ce (

)

Nanog

0

50

100

150

Fold

diff

eren

ce (

)


lowastP lt 005


(b)

EPCs

00

01

02

03

04

05MSCs

00

02

04

06

08

perip

hera

l blo

od

perip

hera

l blo

od



Num

ber o

f EPC

s1120583

L

Num

ber o

f MSC

s1120583

L

(c)





4 Discussion












Acknowledgments


References












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




4 Discussion












Acknowledgments


References












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology





Acknowledgments


References












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology





















Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Endurance Exercise Mobilizes Developmentally Early Stem Cells into Peripheral Blood and Increases...

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Transcript of Endurance Exercise Mobilizes Developmentally Early Stem Cells into Peripheral Blood and Increases...