Cardiomyocyte differentiation of perinatally‑derived mesenchymal stem cells

Nartprayut,Kuneerat; U‑Pratya,Yaowalak; Kheolamai,Pakpoom; Manochantr,Sirikul; Chayosumrit,Methichit; Issaragrisil,Surapol; Supokawej,Aungkura

doi:10.3892/mmr.2013.1356

Cardiomyocyte differentiation of perinatally‑derived mesenchymal stem cells

Authors:
- Kuneerat Nartprayut
- Yaowalak U‑Pratya
- Pakpoom Kheolamai
- Sirikul Manochantr
- Methichit Chayosumrit
- Surapol Issaragrisil
- Aungkura Supokawej
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Affiliations: Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand, Division of Hematology, Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand, Division of Cell Biology, Department of Pre‑Clinical Sciences, Faculty of Medicine, Thammasat University, Pathumthani, Thailand, Siriraj Center of Excellence for Stem Cell Research, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand
Published online on: March 4, 2013 https://doi.org/10.3892/mmr.2013.1356
Pages: 1465-1469

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Abstract

Coronary heart disease is major cause of mortality worldwide and several risk factors have been shown to play a role in its pathogenesis, including smoking, obesity, hypertension and hypercholesterolemia. A number of therapeutic methods have been developed to improve the quality of patients' lives, including stem cell therapy using mesenchymal stem cells (MSCs). Perinatal sources, including the placenta (PL) and umbilical cord (UC), are rich sources of MSCs and have been identified as a potential source of cells for therapeutic use. Their role in cardiogenic differentiation is also of contemporary medical interest. The present study demonstrated the induced differentiation of MSCs obtained from the UC, PL and Wharton's jelly (WJ) into cardiomyocytes, using 10 µM 5‑azacytidine. The characteristics of the MSCs from each source were studied and their morphology was compared. An immunofluorescence analysis for the cardiac‑specific markers, GATA4 and Troponin T (TnT), was performed and tested positive in all sources. The expression of the cardiac‑specific genes, Nkx2.5, α‑cardiac actin and TnT, was analyzed by real‑time RT‑PCR and presented as fold change increases. The expression of each of the markers was observed to be higher in the 5‑azacytidine‑treated MSCs. The differences in expression among the sources of treated MSCs was as follows: TnT had the highest level of expression in the bone marrow (BM) MSCs; α‑cardiac actin had the highest level of expression in the PLMSCs; and all the genes were expressed at significantly high levels in the WJMSCs compared with the control group. The present study showed the ability of alternative perinatally‑derived MSCs to differentiate into cardiomyocyte‑like cells and how this affects the therapeutic use of these cells.

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1	Friedenstein AJ, Chailakhyan RK, Latsinik NV, Panasyuk AF and Keiliss-Borok IV: Stromal cells responsible for transferring the microenvironment of the hemopoietic tissues. Cloning in vitro and retransplantation in vivo. Transplantation. 17:331–340. 1974. View Article : Google Scholar : PubMed/NCBI
2	Pittenger MF, Mackay AM, Beck SC, et al: Multilineage potential of adult human mesenchymal stem cells. Science. 284:143–147. 1999. View Article : Google Scholar : PubMed/NCBI
3	Wu Y, Chen L, Scott PG and Tredget EE: Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells. 25:2648–2659. 2007. View Article : Google Scholar : PubMed/NCBI
4	Morigi M, Introna M, Imberti B, et al: Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice. Stem Cells. 26:2075–2082. 2008. View Article : Google Scholar : PubMed/NCBI
5	Fu X and Li H: Mesenchymal stem cells and skin wound repair and regeneration: possibilities and questions. Cell Tissue Res. 335:317–321. 2009. View Article : Google Scholar : PubMed/NCBI
6	Richardson SM, Hoyland JA, Mobasheri R, Csaki C, Shakibaei M and Mobasheri A: Mesenchymal stem cells in regenerative medicine: opportunities and challenges for articular cartilage and intervertebral disc tissue engineering. J Cell Physiol. 222:23–32. 2010. View Article : Google Scholar : PubMed/NCBI
7	Mueller SM and Glowacki J: Age-related decline in the osteogenic potential of human bone marrow cells cultured in three-dimensional collagen sponges. J Cell Biochem. 82:583–590. 2001. View Article : Google Scholar : PubMed/NCBI
8	Bunnell BA, Flaat M, Gagliardi C, Patel B and Ripoll C: Adipose-derived stem cells: isolation, expansion and differentiation. Methods. 45:115–120. 2008. View Article : Google Scholar : PubMed/NCBI
9	Lee OK, Kuo TK, Chen WM, Lee KD, Hsieh SL and Chen TH: Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood. 103:1669–1675. 2004. View Article : Google Scholar : PubMed/NCBI
10	Villaron EM, Almeida J, López-Holgado N, et al: Mesenchymal stem cells are present in peripheral blood and can engraft after allogeneic hematopoietic stem cell transplantation. Haematologica. 89:1421–1427. 2004.PubMed/NCBI
11	Wang HS, Hung SC, Peng ST, et al: Mesenchymal stem cells in the Wharton’s jelly of the human umbilical cord. Stem Cells. 22:1330–1337. 2004.
12	Surbek D, Wagner A and Schoeberlein A: Perinatal stem cell therapy. Perinatal Stem Cells. Cetrulo CL, Cetrulo KJ and Cetrulo CL Jr: John Wiley & Sons; Hoboken, NJ: pp. 51–60. 2009
13	Dominici M, Le Blanc K, Mueller I, et al: Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 8:315–317. 2006. View Article : Google Scholar
14	Shim WS, Jiang S, Wong P, et al: Ex vivo differentiation of human adult bone marrow stem cells into cardiomyocyte-like cells. Biochem Biophys Res Commun. 324:481–488. 2004. View Article : Google Scholar : PubMed/NCBI
15	Hattan N, Kawaguchi H, ando K, et al: Purified cardiomyocytes from bone marrow mesenchymal stem cells produce stable intracardiac grafts in mice. Cardiovasc Res. 65:334–344. 2005. View Article : Google Scholar : PubMed/NCBI
16	Kruglyakov PV, Sokolova IB, Zin’kova NN, et al: In vitro and in vivo differentiation of mesenchymal stem cells in the cardiomyocyte direction. Bull Exp Biol Med. 142:503–506. 2006. View Article : Google Scholar : PubMed/NCBI
17	Boyum A: Isolation of leucocytes from human blood. A two-phase system for removal of red cells with methylcellulose as erythrocyte-aggregating agent. Scand J Clin Lab Invest Suppl. 97:9–29. 1968.PubMed/NCBI
18	Xu W, Zhang X, Qian H, et al: Mesenchymal stem cells from adult human bone marrow differentiate into a cardiomyocyte phenotype in vitro. Exp Biol Med (Maywood). 229:623–631. 2004.PubMed/NCBI
19	Tsai MS, Hwang SM, Tsai YL, Cheng FC, Lee JL and Chang YJ: Clonal amniotic fluid-derived stem cells express characteristics of both mesenchymal and neural stem cells. Biol Reprod. 74:545–551. 2006. View Article : Google Scholar : PubMed/NCBI
20	Wu KH, Zhou B, Lu SH, et al: In vitro and in vivo differentiation of human umbilical cord derived stem cells into endothelial cells. J Cell Biochem. 100:608–616. 2007. View Article : Google Scholar : PubMed/NCBI
21	Kadivar M, Khatami S, Mortazavi Y, Shokrgozar MA, Taghikhani M and Soleimani M: In vitro cardiomyogenic potential of human umbilical vein-derived mesenchymal stem cells. Biochem Biophys Res Commun. 340:639–647. 2006. View Article : Google Scholar : PubMed/NCBI
22	Benayahu D, Shefer G and Shur I: Insights into the transcriptional and chromatin regulation of mesenchymal stem cells in musculo-skeletal tissues. Ann Anat. 191:2–12. 2009. View Article : Google Scholar : PubMed/NCBI
23	Collas P: Epigenetic states in stem cells. Biochim Biophys Acta. 1790:900–905. 2009. View Article : Google Scholar : PubMed/NCBI
24	Martin-Rendon E, Sweeney D, Lu F, Girdlestone J, Navarrete C and Watt SM: 5-Azacytidine-treated human mesenchymal stem/progenitor cells derived from umbilical cord, cord blood and bone marrow do not generate cardiomyocytes in vitro at high frequencies. Vox Sang. 95:137–148. 2008. View Article : Google Scholar
25	Zhang Y, Chu Y, Shen W and Dou Z: Effect of 5-azacytidine induction duration on differentiation of human first-trimester fetal mesenchymal stem cells towards cardiomyocyte-like cells. Interact Cardiovasc Thorac Surg. 9:943–946. 2009. View Article : Google Scholar
26	Nagaya N, Kangawa K, Itoh T, et al: Transplantation of mesenchymal stem cells improves cardiac function in a rat model of dilated cardiomyopathy. Circulation. 112:1128–1135. 2005. View Article : Google Scholar : PubMed/NCBI
27	Silva GV, Litovsky S, Assad JA, et al: Mesenchymal stem cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a canine chronic ischemia model. Circulation. 111:150–156. 2005. View Article : Google Scholar
28	Shake JG, Gruber PJ, Baumgartner WA, et al: Mesenchymal stem cell implantation in a swine myocardial infarct model: engraftment and functional effects. Ann Thorac Surg. 73:1919–1926. 2002. View Article : Google Scholar : PubMed/NCBI
29	Guo J, Lin GS, Bao CY, Hu ZM and Hu MY: Anti-inflammation role for mesenchymal stem cells transplantation in myocardial infarction. Inflammation. 30:97–104. 2007. View Article : Google Scholar : PubMed/NCBI
30	Markel TA, Wang Y, Herrmann JL, et al: VEGF is critical for stem cell-mediated cardioprotection and a crucial paracrine factor for defining the age threshold in adult and neonatal stem cell function. Am J Physiol Heart Circ Physiol. 295:H2308–H2314. 2008. View Article : Google Scholar : PubMed/NCBI
31	Copland IB and Galipeau J: Death and inflammation following somatic cell transplantation. Semin Immunopathol. 33:535–550. 2011. View Article : Google Scholar : PubMed/NCBI