Tumorigenesis of nuclear transfer-derived embryonic stem cells is reduced through differentiation and enrichment following transplantation in the infarcted rat heart

Fu,Qiang; Su,Dechun; Wang,Ke; Zhao,Yingjun

doi:10.3892/mmr.2016.5092

Tumorigenesis of nuclear transfer-derived embryonic stem cells is reduced through differentiation and enrichment following transplantation in the infarcted rat heart

Authors:
- Qiang Fu
- Dechun Su
- Ke Wang
- Yingjun Zhao
View Affiliations

Affiliations: Department of Cardiology, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China, Department of Cardiology, The People's Hospital of Liaoning Province, Shenyang, Liaoning 110016, P.R. China
Published online on: April 7, 2016 https://doi.org/10.3892/mmr.2016.5092
Pages: 4659-4665

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to evaluate the tumorigenic potential of nuclear transfer-derived (nt) mouse embryonic stem cells (mESCs) transplanted into infarcted rat hearts. The nt‑mESCs were cultured using a bioreactor system to develop embryoid bodies, which were induced with 1% ascorbic acid to differentiate into cardiomyocytes. The nt‑mESC‑derived cardiomyocytes (nt‑mESCs‑CMs) were enriched using Percoll density gradient separation to generate nt‑mESCs‑percoll‑enriched (PE)‑CMs. Ischemia was induced by ligating the left anterior descending coronary artery in female Sprague‑Dawley rats. Immunosuppressed rats (daily intraperitoneal injections of cyclosporine A and methylprednisolone) were randomly assigned to receive an injection containing 5x106 mESCs, nt‑mESCs, nt‑mESC‑CMs or nt‑mESC‑PE‑CMs. Analysis performed 8 weeks following transplantation revealed teratoma formation in 80, 86.67 and 33.33% of the rats administered with the mESCs, nt‑mESCs and nt‑mESC‑CMs, respectively, indicating no significant difference between the mESCs and nt‑mESCs; but significance (P<0.05) between the nt‑mESC‑CMs and nt‑mESCs. The mean tumor volumes were 82.72±6.52, 83.17±3.58 and 50.40±5.98 mm3, respectively (P>0.05 mESCs, vs. nt‑mESCs; P<0.05 nt‑mESC‑CMs, vs. nt‑mESCs). By contrast, no teratoma formation was detected in the rats, which received nt‑mESC‑PE‑CMs. Octamer‑binding transcription factor‑4, a specific marker of undifferentiated mESCs, was detected using polymerase chain reaction in the rats, which received nt‑mESCs and nt‑mESC‑CMs, but not in rats administered with nt‑mESC‑PE‑CMs. In conclusion, nt‑mESCs exhibited the same pluripotency as mESCs, and teratoma formation following nt‑mESC transplantation was reduced by cell differentiation and enrichment.

View Figures

View References

1	Soonpaa MH and Field LJ: Survey of studies examining mammalian cardiomyocyte DNA synthesis. Circ Res. 83:15–26. 1998. View Article : Google Scholar : PubMed/NCBI
2	Sun Y and Weber KT: Infarct scar: A dynamic tissue. Cardiovasc Res. 46:250–256. 2000. View Article : Google Scholar : PubMed/NCBI
3	Taylor DO, Edwards LB, Aurora P, Christie JD, Dobbels F, Kirk R, Rahmel AO, Kucheryavaya AY and Hertz MI: Registry of the international society for heart and lung transplantation: Twenty-fifth official adult heart transplant report-2008. J Heart Lung Transplant. 27:943–956. 2008. View Article : Google Scholar : PubMed/NCBI
4	Augoustides JG and Riha H: Recent progress in heart failure treatment and heart transplantation. J Cardiothorac Vasc Anesth. 23:738–748. 2009. View Article : Google Scholar : PubMed/NCBI
5	Laflamme MA and Murry CE: Regenerating the heart. Nat Biotechnol. 23:845–856. 2005. View Article : Google Scholar : PubMed/NCBI
6	Laflamme MA, Zbinden S, Epstein SE and Murry CE: Cell-based therapy for myocardial ischemia and infarction: Pathophysiological mechanisms. Annu Rev Pathol. 2:307–339. 2007. View Article : Google Scholar : PubMed/NCBI
7	Soonpaa MH, Koh GY, Klug MG and Field LJ: Formation of nascent intercalated disks between grafted fetal cardiomyocytes and host myocardium. Science. 264:98–101. 1994. View Article : Google Scholar : PubMed/NCBI
8	Reinecke H, Zhang M, Bartosek T and Murry CE: Survival, integration, and differentiation of cardiomyocyte grafts: A study in normal and injured rat hearts. Circulation. 100:193–202. 1999. View Article : Google Scholar : PubMed/NCBI
9	Koh GY, Soonpaa MH, Klug MG, Pride HP, Cooper BJ, Zipes DP and Field LJ: Stable fetal cardiomyocyte grafts in the hearts of dystrophic mice and dogs. J Clin Invest. 96:2034–2042. 1995. View Article : Google Scholar : PubMed/NCBI
10	Evans MJ and Kaufman MH: Establishment in culture of pluripotential cells from mouse embryos. Nature. 292:154–156. 1981. View Article : Google Scholar : PubMed/NCBI
11	Amit M, Carpenter MK, Inokuma MS, Chiu CP, Harris CP, Waknitz MA, Itskovitz-Eldor J and Thomson JA: Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. Dev Biol. 227:271–278. 2000. View Article : Google Scholar : PubMed/NCBI
12	Xu C, Police S, Rao N and Carpenter MK: Characterization and enrichment of cardiomyocytes derived from human embryonic stem cells. Circ Res. 91:501–508. 2002. View Article : Google Scholar : PubMed/NCBI
13	Mummery C, Ward-van Oostwaard D, Doevendans P, Spijker R, van den Brink S, Hassink R, van der Heyden M, Opthof T, Pera M, de la Riviere AB, et al: Differentiation of human embryonic stem cells to cardiomyocytes: Role of coculture with visceral endoderm-like cells. Circulation. 107:2733–2740. 2003. View Article : Google Scholar : PubMed/NCBI
14	Xie CQ, Zhang J, Xiao Y, Zhang L, Mou Y, Liu X, Akinbami M, Cui T and Chen YE: Transplantation of human undifferentiated embryonic stem cells into a myocardial infarction rat model. Stem Cells Dev. 16:25–29. 2007. View Article : Google Scholar : PubMed/NCBI
15	Caspi O, Huber I, Kehat I, Habib M, Arbel G, Gepstein A, Yankelson L, Aronson D, Beyar R and Gepstein L: Transplantation of human embryonic stem cell-derived cardiomyocytes improves myocardial performance in infarcted rat hearts. J Am Coll Cardiol. 50:1884–1893. 2007. View Article : Google Scholar : PubMed/NCBI
16	Robbins J, Gulick J, Sanchez A, Howles P and Doetschman T: Mouse embryonic stem cells express the cardiac myosin heavy chain genes during development in vitro. J Biol Chem. 265:11905–11909. 1990.PubMed/NCBI
17	Maltsev VA, Wobus AM, Rohwedel J, Bader M and Hescheler J: Cardiomyocytes differentiated in vitro from embryonic stem cells developmentally express cardiacspecific genes and ionic currents. Circ Res. 75:233–244. 1994. View Article : Google Scholar : PubMed/NCBI
18	Yang Y, Min JY, Rana JS, Ke Q, Cai J, Chen Y, Morgan JP and Xiao YF: VEGF enhances functional improvement of postin-farcted hearts by transplantation of ESC-differentiated cells. J Appl Physiol. 93:1140–1151. 2002. View Article : Google Scholar
19	Laflamme MA, Chen KY, Naumova AV, Muskheli V, Fugate JA, Dupras SK, Reinecke H, Xu C, Hassanipour M, Police S, et al: Cardiomyocytes derived from human embryonic stem cells in pro-survival factors enhance function of infarcted rat hearts. Nat Biotechnol. 25:1015–1024. 2007. View Article : Google Scholar : PubMed/NCBI
20	van Laake LW, Passier R, Monshouwer-Kloots J, Verkleij AJ, Lips DJ, Freund C, den Ouden K, Ward-van Oostwaard D, Korving J, Tertoolen LG, et al: Human embryonic stem cell-derived cardiomyocytes survive and mature in the mouse heart and transiently improve function after myocardial infarction. Stem Cell Res. 1:9–24. 2007. View Article : Google Scholar : PubMed/NCBI
21	Swijnenburg RJ, Tanaka M, Vogel H, Baker J, Kofidis T, Gunawan F, Lebl DR, Caffarelli AD, de Bruin JL, Fedoseyeva EV and Robbins RC: Embryonic stem cell immunogenicity increases upon differentiation after transplantation into ischemic myocardium. Circulation. 112(9 Suppl): I166–I172. 2005.PubMed/NCBI
22	He Q, Trindade PT, Stumm M, Li J, Zammaretti P, Bettiol E, Dubois-Dauphin M, Herrmann F, Kalangos A, Morel D, et al: Fate of undifferentiated mouse embryonic stem cells within the rat heart: Role of myocardial infarction and immune suppression. J Cell Mol Med. 13:188–201. 2009. View Article : Google Scholar
23	Nussbaum J, Minami E, Laflamme MA, Virag JA, Ware CB, Masino A, Muskheli V, Pabon L, Reinecke H and Murry CE: Transplantation of undifferentiated murine embryonic stem cells in the heart:Teratoma formation and immune response. Fasbe J. 21:1345–1357. 2007. View Article : Google Scholar
24	Willadsen SM: Nuclear transplantation in sheep embryos. Nature. 320:63–65. 1986. View Article : Google Scholar : PubMed/NCBI
25	Cibelli JB, Stice SL, Golueke PJ, et al: Transgenic bovine chimeric offspring produced from somatic cell-derived stem-like cells. Nat Biotechnol. 16:642–646. 1998. View Article : Google Scholar : PubMed/NCBI
26	Koh CJ and Atala A: Tissue engineering, stem cells, and cloning: Opportunities for regenerative medicine. J Am Soc Nephrol. 15:1113–1125. 2004. View Article : Google Scholar : PubMed/NCBI
27	Leor J, Gerecht S, Cohen S, Miller L, Holbova R, Ziskind A, Shachar M, Feinberg MS, Guetta E and Itskovitz-Eldor J: Human embryonic stem cell transplantation to repair the infarcted myocardium. Heart. 93:1278–1284. 2007. View Article : Google Scholar : PubMed/NCBI
28	Cooke MJ, Stojkovic M and Przyborski SA: Growth of teratomas derived from human pluripotent stem cells is influenced by the graft site. Stem Cells Dev. 15:254–259. 2006. View Article : Google Scholar : PubMed/NCBI
29	Prokhorova TA, Harkness LM, Frandsen U, Ditzel N, Schrøder HD, Burns JS and Kassem M: Teratoma formation by human embryonic stem cells is site dependent and enhanced by the presence of Matrigel. Stem Cells Dev. 18:47–54. 2009. View Article : Google Scholar
30	Toumadje A, Kusumoto K, Parton A, Mericko P, Dowell L, Ma G, Chen L, Barnes DW and Sato JD: Pluripotent differentiation in vitro of murine ES-D3 embryonic stem cells. In Vitro Cell Dev Biol Anim. 39:449–453. 2003. View Article : Google Scholar
31	Miyahara Y, Nagaya N, Kataoka M, Yanagawa B, Tanaka K, Hao H, Ishino K, Ishida H, Shimizu T, Kangawa K, et al: Monolayered mesenchymal stem cells repair scarred myocardium after myocardial infarction. Nat Med. 12:459–465. 2006. View Article : Google Scholar : PubMed/NCBI
32	Lin Q, Fu Q, Zhang Y, Wang H, Liu Z, Zhou J, Duan C, Wang Y, Wu K and Wang C: Tumourigenesis in the infarcted rat heart is eliminated through differentiation and enrichment of the transplanted embryonic stem cells. Eur J Heart Fail. 12:1179–1185. 2010. View Article : Google Scholar : PubMed/NCBI
33	Reubinoff BE, Pera MF, Fong CY, Trounson A and Bongso A: Embryonic stem cell lines from human blastocysts: Somatic differentiation in vitro. Nat Biotechnol. 18:399–404. 2000. View Article : Google Scholar : PubMed/NCBI
34	Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS and Jones JM: Embryonic stem cell lines derived from human blastocysts. Science. 282:1145–1147. 1998. View Article : Google Scholar : PubMed/NCBI
35	Laflamme MA, Gold J, Xu C, Hassanipour M, Rosler E, Police S, Muskheli V and Murry CE: Formation of human myocardium in the rat heart from human embryonic stem cells. Am J Pathol. 167:663–671. 2005. View Article : Google Scholar : PubMed/NCBI