Cell therapy of heart diseases is emerging as one of the most promising known treatments in recent years. Transplantation of cardiac stem cells (CSCs) may be one of the best strategies to cure adult or pediatric heart diseases. As these patient-derived stem cells need to be isolated from small heart biopsies, it is important to select the best isolation method and CSC subpopulation with the best cardiogenic functionality. We employed three different protocols including c-KIT+ cell sorting, clonogenic expansion, and explants culture to isolate c-KIT+ cells, clonogenic expansion-derived cells (CEDCs), and cardiosphere-derived cells (CDCs), respectively. Evaluation of isolated CSC characteristics in vitro and after rat myocardial infarction (MI) model transplantation revealed that although c-KIT+ and CDCs had higher MI regenerative potential, CEDCs had more commitment into cardiomyocytes and needed lower passages that were essential to reach a definite cell count. Furthermore, genome-wide expression analysis showed that subsequent passages caused changes in characteristics of cells, downregulation of cell cycle-related genes, and upregulation of differentiation and carcinogenic genes, which might lead to senescence, commitment, and possible tumorigenicity of the cells. Because of different properties of CSC subpopulations, we suggest that appropriate CSCs subpopulation should be chosen based on their experimental or clinical use.
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References
1.
LozanoR, NaghaviM, ForemanK, LimS, ShibuyaK, AboyansV, AbrahamJ, AdairT, AggarwalRet al., (2012). Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet, 380:2095–2128.
2.
FinegoldJA, AsariaP and FrancisDP. (2013). Mortality from ischaemic heart disease by country, region, and age: statistics from World Health Organisation and United Nations. Int J Cardiol, 168:934–945.
3.
SmithRR, BarileL, MessinaE and MarbanE. (2008). Stem cells in the heart: what's the buzz all about?. Part 2: arrhythmic risks and clinical studies. Heart Rhythm, 5:880–887.
4.
FilesMD and BoucekRJ. (2012). ‘Shovel-Ready’ applications of stem cell advances for pediatric heart disease. Curr Opin Pediatr, 24:577–583.
5.
MakkarRR, SmithRR, ChengK, MalliarasK, ThomsonLE, BermanD, CzerLS, MarbanL, MendizabalAet al., (2012). Intracoronary cardiosphere-derived cells for heart regeneration after myocardial infarction (CADUCEUS): a prospective, randomised phase 1 trial. Lancet, 379:895–904.
6.
BolliR, ChughAR, D'AmarioD, LoughranJH, StoddardMF, IkramS, BeacheGM, WagnerSG, LeriAet al., (2011). Cardiac stem cells in patients with ischaemic cardiomyopathy (SCIPIO): initial results of a randomised phase 1 trial. Lancet, 378:1847–1857.
7.
ZhaoX and HuangL. (2013). Cardiac stem cells: a promising treatment option for heart failure. Exp Ther Med, 5:379–383.
8.
MishraR, VijayanK, CollettiEJ, HarringtonDA, MatthiesenTS, SimpsonD, GohSK, WalkerBL, Almeida-PoradaGet al., (2011). Characterization and functionality of cardiac progenitor cells in congenital heart patients. Circulation, 123:364–373.
9.
SmithRR, BarileL, MessinaE and MarbanE. (2008). Stem cells in the heart: what's the buzz all about?.—Part 1: preclinical considerations. Heart Rhythm, 5:749–757.
10.
BeltramiAP, BarlucchiL, TorellaD, BakerM, LimanaF, ChimentiS, KasaharaH, RotaM, MussoEet al., (2003). Adult cardiac stem cells are multipotent and support myocardial regeneration. Cell, 114:763–776.
11.
OhH, BradfuteSB, GallardoTD, NakamuraT, GaussinV, MishinaY, PociusJ, MichaelLH, BehringerRRet al., (2003). Cardiac progenitor cells from adult myocardium: homing, differentiation, and fusion after infarction. Proc Natl Acad Sci U S A, 100:12313–12318.
12.
MessinaE, De AngelisL, FratiG, MorroneS, ChimentiS, FiordalisoF, SalioM, BattagliaM, LatronicoMVet al., (2004). Isolation and expansion of adult cardiac stem cells from human and murine heart. Circ Res, 95:911–921.
13.
BearziC, RotaM, HosodaT, TillmannsJ, NascimbeneA, De AngelisA, Yasuzawa-AmanoS, TrofimovaI, SigginsRWet al., (2007). Human cardiac stem cells. Proc Natl Acad Sci U S A, 104:14068–14073.
14.
ZarubaMM, SoonpaaM, ReuterS and FieldLJ. (2010). Cardiomyogenic potential of C-kit(+)-expressing cells derived from neonatal and adult mouse hearts. Circulation, 121:1992–2000.
15.
D'AmarioD, FioriniC, CampbellPM, GoichbergP, SanadaF, ZhengH, HosodaT, RotaM, ConnellJMet al., (2011). Functionally competent cardiac stem cells can be isolated from endomyocardial biopsies of patients with advanced cardiomyopathies. Circ Res, 108:857–861.
16.
TangYL, ShenL, QianK and PhillipsMI. (2007). A novel two-step procedure to expand cardiac Sca-1+ cells clonally. Biochem Biophys Res Commun, 359:877–883.
17.
DavisDR, KizanaE, TerrovitisJ, BarthAS, ZhangY, SmithRR, MiakeJ and MarbanE. (2010). Isolation and expansion of functionally-competent cardiac progenitor cells directly from heart biopsies. J Mol Cell Cardiol, 49:312–321.
18.
CarrCA, StuckeyDJ, TanJJ, TanSC, GomesRS, CamellitiP, MessinaE, GiacomelloA, EllisonGM and ClarkeK. (2011). Cardiosphere-derived cells improve function in the infarcted rat heart for at least 16 weeks—an MRI study. PLoS One, 6:e25669.
19.
YeJ, BoyleA, ShihH, SieversRE, ZhangY, PrasadM, SuH, ZhouY, GrossmanW, BernsteinHS and YeghiazariansY. (2012). Sca-1+ cardiosphere-derived cells are enriched for Isl1-expressing cardiac precursors and improve cardiac function after myocardial injury. PLoS One, 7:e30329.
20.
SmitsAM, van VlietP, MetzCH, KorfageT, SluijterJP, DoevendansPA and GoumansMJ. (2009). Human cardiomyocyte progenitor cells differentiate into functional mature cardiomyocytes: an in vitro model for studying human cardiac physiology and pathophysiology. Nat Protoc, 4:232–243.
21.
EllisonGM, Nadal-GinardB and TorellaD. (2012). Optimizing cardiac repair and regeneration through activation of the endogenous cardiac stem cell compartment. J Cardiovasc Transl Res, 5:667–677.
22.
WilsonA, ShehadehLA, YuH and WebsterKA. (2010). Age-related molecular genetic changes of murine bone marrow mesenchymal stem cells. BMC Genomics, 11:229.
23.
LeeMY, WangY and VanhouttePM. (2010). Senescence of cultured porcine coronary arterial endothelial cells is associated with accelerated oxidative stress and activation of NFkB. J Vasc Res, 47:287–298.
24.
ApitzC, WebbGD and RedingtonAN. (2009). Tetralogy of Fallot. Lancet, 374:1462–1471.
LeriA, KajsturaJ and AnversaP. (2005). Cardiac stem cells and mechanisms of myocardial regeneration. Physiol Rev, 85:1373–1416.
31.
TakamiyaM, HaiderKH and AshrafM. (2011). Identification and characterization of a novel multipotent sub-population of Sca-1(+) cardiac progenitor cells for myocardial regeneration. PLoS One, 6:e25265.
32.
Di NardoP, ForteG, AhluwaliaA and MinieriM. (2010). Cardiac progenitor cells: potency and control. J Cell Physiol, 224:590–600.
33.
LiTS, ChengK, LeeST, MatsushitaS, DavisD, MalliarasK, ZhangY, MatsushitaN, SmithRR and MarbanE. (2010). Cardiospheres recapitulate a niche-like microenvironment rich in stemness and cell-matrix interactions, rationalizing their enhanced functional potency for myocardial repair. Stem Cells, 28:2088–2098.
ChimentiI, SmithRR, LiTS, GerstenblithG, MessinaE, GiacomelloA and MarbanE. (2010). Relative roles of direct regeneration versus paracrine effects of human cardiosphere-derived cells transplanted into infarcted mice. Circ Res, 106:971–980.
36.
TangXL, RokoshG, SanganalmathSK, YuanF, SatoH, MuJ, DaiS, LiC, ChenNet al., (2010). Intracoronary administration of cardiac progenitor cells alleviates left ventricular dysfunction in rats with a 30-day-old infarction. Circulation, 121:293–305.
37.
LiTS, ChengK, MalliarasK, SmithRR, ZhangY, SunB, MatsushitaN, BlusztajnA, TerrovitisJet al., (2012). Direct comparison of different stem cell types and subpopulations reveals superior paracrine potency and myocardial repair efficacy with cardiosphere-derived cells. J Am Coll Cardiol, 59:942–953.
38.
SmithRR, BarileL, ChoHC, LeppoMK, HareJM, MessinaE, GiacomelloA, AbrahamMR and MarbanE. (2007). Regenerative potential of cardiosphere-derived cells expanded from percutaneous endomyocardial biopsy specimens. Circulation, 115:896–908.
39.
HuangC, GuH, YuQ, ManukyanMC, PoynterJA and WangM. (2011). Sca-1+ cardiac stem cells mediate acute cardioprotection via paracrine factor SDF-1 following myocardial ischemia/reperfusion. PLoS One, 6:e29246.
40.
FuentesT and Kearns-JonkerM. (2013). Endogenous cardiac stem cells for the treatment of heart failure. Stem Cells Cloning, 6:1–12.
41.
Nadal-GinardB, EllisonGM and TorellaD. (2014). The cardiac stem cell compartment is indispensable for myocardial cell homeostasis, repair and regeneration in the adult. Stem Cell Res, 13:615–630.
42.
den HaanMC, GraussRW, SmitsAM, WinterEM, van TuynJ, PijnappelsDA, SteendijkP, Gittenberger-De GrootAC, van der LaarseAet al., (2012). Cardiomyogenic differentiation-independent improvement of cardiac function by human cardiomyocyte progenitor cell injection in ischaemic mouse hearts. J Cell Mol Med, 16:1508–1521.
43.
YeL, ChangYH, XiongQ, ZhangP, ZhangL, SomasundaramP, LepleyM, SwingenC, SuLet al., (2014). Cardiac repair in a porcine model of acute myocardial infarction with human induced pluripotent stem cell-derived cardiovascular cells. Cell Stem Cell, 15:750–761.
44.
DeyD, HanL, BauerM, SanadaF, OikonomopoulosA, HosodaT, UnnoK, De AlmeidaP, LeriA and WuJC. (2013). Dissecting the molecular relationship among various cardiogenic progenitor cells. Circ Res, 112:1253–1262.
45.
BabuGJ, WarshawDM and PeriasamyM. (2000). Smooth muscle myosin heavy chain isoforms and their role in muscle physiology. Microsc Res Tech, 50:532–540.
46.
BarileL, ChimentiI, GaetaniR, ForteE, MiraldiF, FratiG, MessinaE and GiacomelloA. (2007). Cardiac stem cells: isolation, expansion and experimental use for myocardial regeneration. Nat Clin Pract Cardiovasc Med, 4Suppl 1:S9–S14.
47.
PartovianC and SimonsM. (2008). Stem cell therapies in cardiovascular disease A “realistic”. appraisal. Drug Discov Today Ther Strateg, 5:73–78.
48.
ZhuW, ShiojimaI, ItoY, LiZ, IkedaH, YoshidaM, NaitoAT, NishiJ, UenoHet al., (2008). IGFBP-4 is an inhibitor of canonical Wnt signalling required for cardiogenesis. Nature, 454:345–349.
49.
LiH, ZuoS, PashaZ, YuB, HeZ, WangY, YangX, AshrafM and XuM. (2011). GATA-4 promotes myocardial transdifferentiation of mesenchymal stromal cells via up-regulating IGFBP-4. Cytotherapy, 13:1057–1065.
50.
OikonomopoulosA, SeretiKI, ConyersF, BauerM, LiaoA, GuanJ, CrappsD, HanJK, DongHet al., (2011). Wnt signaling exerts an antiproliferative effect on adult cardiac progenitor cells through IGFBP3. Circ Res, 109:1363–1374.
51.
D'AmarioD, Cabral-Da-SilvaMC, ZhengH, FioriniC, GoichbergP, SteadmanE, Ferreira-MartinsJ, SanadaF, PiccoliMet al., (2011). Insulin-like growth factor-1 receptor identifies a pool of human cardiac stem cells with superior therapeutic potential for myocardial regeneration. Circ Res, 108:1467–1481.
52.
HsiehD, HsiehA, SteaB and EllsworthR. (2010). IGFBP2 promotes glioma tumor stem cell expansion and survival. Biochem Biophys Res Commun, 397:367–372.
53.
ZebedeeZ and HaraE. (2001). Id proteins in cell cycle control and cellular senescence. Oncogene, 20:8317–8325.
54.
YokotaY and MoriS. (2002). Role of Id family proteins in growth control. J Cell Physiol, 190:21–28.
55.
MengQ, JiaZ, WangW, LiB, MaK and ZhouC. (2011). Inhibitor of DNA binding 1 (Id1) induces differentiation and proliferation of mouse embryonic carcinoma P19CL6 cells. Biochem Biophys Res Commun, 412:253–259.
56.
ZhaoQ, BeckAJ, VitaleJM, SchneiderJS, GaoS, ChangC, ElsonG, LeibovichSJ, ParkJYet al., (2011). Developmental ablation of Id1 and Id3 genes in the vasculature leads to postnatal cardiac phenotypes. Dev Biol, 349:53–64.
57.
ImanishiT, HanoT, SawamuraT, TakaradaS and NishioI. (2002). Oxidized low density lipoprotein potentiation of Fas-induced apoptosis through lectin-like oxidized-low density lipoprotein receptor-1 in human umbilical vascular endothelial cells. Circ J, 66:1060–1064.
58.
KoremS, ResnickMB and KraiemZ. (1999). Similar and divergent patterns in the regulation of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of MMP-1 gene expression in benign and malignant human thyroid cells. J Clin Endocrinol Metab, 84:3322–3327.
59.
McCawleyLJ and MatrisianLM. (2000). Matrix metalloproteinases: multifunctional contributors to tumor progression. Mol Med Today, 6:149–156.
60.
StamenkovicI. (2003). Extracellular matrix remodelling: the role of matrix metalloproteinases. J Pathol, 200:448–464.
61.
KimHE, DalalSS, YoungE, LegatoMJ, WeisfeldtML and D'ArmientoJ. (2000). Disruption of the myocardial extracellular matrix leads to cardiac dysfunction. J Clin Invest, 106:857–866.
62.
SappinoAP, HuarteJ, BelinD and VassalliJD. (1989). Plasminogen activators in tissue remodeling and invasion: mRNA localization in mouse ovaries and implanting embryos. J Cell Biol, 109:2471–2479.
63.
McMahonB and KwaanHC. (2008). The plasminogen activator system and cancer. Pathophysiol Haemost Thromb, 36:184–194.
64.
YoshizawaK, NozakiS, KitaharaH, KatoK, NoguchiN, KawashiriS and YamamotoE. (2011). Expression of urokinase-type plasminogen activator/urokinase-type plasminogen activator receptor and maspin in oral squamous cell carcinoma: Association with mode of invasion and clinicopathological factors. Oncol Rep, 26:1555–1560.
65.
ThummaratiP, WijitburaphatS, PrasopthumA, MenakongkaA, SripaB, TohtongR and SuthiphongchaiT. (2012). High level of urokinase plasminogen activator contributes to cholangiocarcinoma invasion and metastasis. World J Gastroenterol, 18:244–250.
66.
DufourcqP, LerouxL, EzanJ, DescampsB, LamaziereJM, CostetP, BasoniC, MoreauC, DeutschU, CouffinhalT and DuplaaC. (2008). Regulation of endothelial cell cytoskeletal reorganization by a secreted frizzled-related protein-1 and frizzled 4- and frizzled 7-dependent pathway: role in neovessel formation. Am J Pathol, 172:37–49.
67.
BergmannMW. (2010). WNT signaling in adult cardiac hypertrophy and remodeling: lessons learned from cardiac development. Circ Res, 107:1198–1208.
68.
SilkeJ and VauxDL. (2001). Two kinds of BIR-containing protein—inhibitors of apoptosis, or required for mitosis. J Cell Sci, 114:1821–1827.
69.
LamersF, van der PloegI, SchildL, EbusME, KosterJ, HansenBR, KochT, VersteegR, CaronHN and MolenaarJJ. (2011). Knockdown of survivin (BIRC5) causes apoptosis in neuroblastoma via mitotic catastrophe. Endocr Relat Cancer, 18:657–668.
70.
MitaAC, MitaMM, NawrockiST and GilesFJ. (2008). Survivin: key regulator of mitosis and apoptosis and novel target for cancer therapeutics. Clin Cancer Res, 14:5000–5005.
71.
LogarinhoE and BousbaaH. (2008). Kinetochore-microtubule interactions “in check”. by Bub1, Bub3 and BubR1: The dual task of attaching and signalling. Cell Cycle, 7:1763–1768.
72.
Bolanos-GarciaVM and BlundellTL. (2011). BUB1 and BUBR1: multifaceted kinases of the cell cycle. Trends Biochem Sci, 36:141–150.
73.
YaoY and DaiW. (2012). Mitotic checkpoint control and chromatin remodeling. Front Biosci, 17:976–983.
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