Tree shrew umbilical cord mesenchymal stem cells: isolation, cultivation and osteogenic and adipogenic differentiation

doi:10.3969/j.issn.2095-4344.2017.09.012

Abstract

Abstract:

BACKGROUND: Studies have shown that umbilical cord mesenchymal stem cells are ideal seed cells for tissue engineering research.
OBJECTIVE: To isolate, culture and identify tree shrew umbilical cord mesenchymal stem cells, in order to establish a standardized tree shrew umbilical cord mesenchymal stem cell lines.
METHODS: Caesarean-isolated tree shrew umbilical cord samples were used to isolate and culture umbilical cord mesenchymal stem cells using tissue explant adherent method. Flow cytometry assay was used to detect cell surface markers. Osteogenic and adipogenic induction media were used to induce umbilical cord mesenchymal stem cells to differentiate into osteoblasts and adipocytes.
RESULTS AND CONCLUSION: The cultured umbilical cord mesenchymal stem cells expressed CD90 and CD105 with the positive rate of 99.9% and 99.8% respectively. Hematopoietic stem cell marker CD34 expression rate was 0.0% and the endothelial cell marker CD31 expression rate was 0.7%, in line with the characteristics of umbilical cord mesenchymal stem cell surface markers. Calcium nodules by alizarin red staining and lipid droplets by oil red O staining were observed in the induced cells. These experimental findings indicated that umbilical cord mesenchymal stem cells from tree shrews capable of osteogenic and adipogenic differentiation were successfully isolated and cultured.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Cell Differentiation, Tupaiidae, Tissue Engineering

CLC Number:

R394.2

Ruan Guang-ping, Zhu Lu, Liu Ju-fen, Li Zi-an, Wang Jin-xiang, Pang Rong-qing, Pan Xing-hua. Tree shrew umbilical cord mesenchymal stem cells: isolation, cultivation and osteogenic and adipogenic differentiation[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(9): 1373-1377.

Figures/Tables 1

References

[1] Barry FP, Murphy JM. Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol. 2004;36(4):568-584.

[2] Mok PL, Leong CF, Cheong SK. Cellular mechanisms of emerging applications of mesenchymal stem cells. Malays J Pathol. 2013;35(1):17-32.

[3] Neirinckx V, Coste C, Rogister B, et al. Concise review: adult mesenchymal stem cells, adult neural crest stem cells, and therapy of neurological pathologies: a state of play. Stem Cells Transl Med. 2013;2(4):284-296.

[4] Van Pham P, Truong NC, Le PT, et al. Isolation and proliferation of umbilical cord tissue derived mesenchymal stem cells for clinical applications. Cell Tissue Bank. 2016; 17(2):289-302.

[5] Lian J, Lv S, Liu C, et al. Effects of serial passage on the characteristics and cardiac and neural differentiation of human umbilical cord wharton's jelly-derived mesenchymal stem cells. Stem Cells Int. 2016;2016:9291013.

[6] Gokcinar-Yagci B, Ozyuncu O, Celebi-Saltik B. Isolation, characterisation and comparative analysis of human umbilical cord vein perivascular cells and cord blood mesenchymal stem cells. Cell Tissue Bank. 2016;17(2):345-352.

[7] Venugopal P, Balasubramanian S, Majumdar AS, et al. Isolation, characterization, and gene expression analysis of Wharton's jelly-derived mesenchymal stem cells under xeno-free culture conditions. Stem Cells Cloning. 2011;4:39-50.

[8] Kim DW, Staples M, Shinozuka K, et al. Wharton's jelly-derived mesenchymal stem cells: phenotypic characterization and optimizing their therapeutic potential for clinical applications. Int J Mol Sci. 2013;14(6):11692-11712.

[9] Banitalebi Dehkordi M, Madjd Z, Chaleshtori MH, et al. A Simple, rapid, and efficient method for isolating mesenchymal stem cells from the entire umbilical cord. Cell Transplant. 2016;25(7):1287-1297.

[10] Ruan ZB, Zhu L, Yin YG, et al. Inhibitor of p53-p21 pathway induces the differentiation of human umbilical cord derived mesenchymal stem cells into cardiomyogenic cells. Cytotechnology. 2016;68(4):1257-1265.

[11] Gong X, Wang P, Wu Q, et al. Human umbilical cord blood derived mesenchymal stem cells improve cardiac function in cTnT(R141W) transgenic mouse of dilated cardiomyopathy. Eur J Cell Biol. 2016;95(1):57-67.

[12] Wang Y, Ying Y, Cui X. Effects on proliferation and differentiation of human umbilical cord-derived mesenchymal stem cells engineered to express neurotrophic factors. Stem Cells Int. 2016;2016:1801340.

[13] Kim SW, Jin HL, Kang SM, et al. Therapeutic effects of late outgrowth endothelial progenitor cells or mesenchymal stem cells derived from human umbilical cord blood on infarct repair. Int J Cardiol. 2016;203:498-507.

[14] Cao H, Hui Q, Yan Y, et al. Pretreatments with injured microenvironmental signals altered the characteristics of human umbilical cord mesenchymal stem cells. Biotechnol Lett. 2016;38(1):157-165.

[15] Xie Z, Hao H, Tong C, et al. Human umbilical cord-derived mesenchymal stem cells elicit macrophages into an anti-inflammatory phenotype to alleviate insulin resistance in type 2 diabetic rats. Stem Cells, 2016;34(3):627-639.

[16] Ren S, Hu J, Chen Y, et al. Human umbilical cord derived mesenchymal stem cells promote interleukin-17 production from human peripheral blood mononuclear cells of healthy donors and systemic lupus erythematosus patients. Clin Exp Immunol. 2016;183(3):389-396.

[17] Montanucci P, Alunno A, Basta G, et al. Restoration of t cell substes of patients with type 1 diabetes mellitus by microencapsulated human umbilical cord Wharton jelly-derived mesenchymal stem cells: an in vitro study. Clin Immunol. 2016;163:34-41.

[18] Fang B, Shi M, Liao L, et al. Multiorgan engraftment and multilineage differentiation by human fetal bone marrow Flk1+/CD31-/CD34- Progenitors. J Hematother Stem Cell Res. 2003;12(6):603-613.

[19] Fang B, Liao L, Shi M, et al. Multipotency of Flk1CD34 progenitors derived from human fetal bone marrow. J Lab Clin Med. 2004;143(4):230-240.

[20] Jiang Y, Jahagirdar BN, Reinhardt RL, et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature. 2002;418(6893):41-49.

[21] Schwartz RE, Reyes M, Koodie L, et al. Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells. J Clin Invest. 2002;109(10):1291-302.

[22] Wang S, Yang H, Tang Z, et al. Wound dressing model of human umbilical cord mesenchymal stem cells-alginates complex promotes skin wound healing by paracrine signaling. Stem Cells Int. 2016;2016:3269267.

[23] Heo JS, Choi Y, Kim HS, et al. Comparison of molecular profiles of human mesenchymal stem cells derived from bone marrow, umbilical cord blood, placenta and adipose tissue. Int J Mol Med. 2016;37(1):115-25.

[24] Hayashi K, Ohta H, Kurimoto K, et al. Reconstitution of the mouse germ cell specification pathway in culture by pluripotent stem cells. Cell. 2011;146(4):519-532.

[25] Bai J, Hu Y, Wang YR, et al. Comparison of human amniotic fluid-derived and umbilical cord Wharton's Jelly-derived mesenchymal stromal cells: characterization and myocardial differentiation capacity. J Geriatr Cardiol. 2012;9(2):166-171.

[26] Xu Y, Meng H, Li C, et al. Umbilical cord-derived mesenchymal stem cells isolated by a novel explantation technique can differentiate into functional endothelial cells and promote revascularization. Stem Cells Dev. 2010;19(10): 1511-1522.

[27] Chen Y, Qian H, Zhu W, et al. Hepatocyte growth factor modification promotes the amelioration effects of human umbilical cord mesenchymal stem cells on rat acute kidney injury. Stem Cells Dev. 2011;20(1):103-113.

[28] Ribeiro J, Pereira T, Amorim I, et al. Cell therapy with human MSCs isolated from the umbilical cord Wharton jelly associated to a PVA membrane in the treatment of chronic skin wounds. Int J Med Sci. 2014;11(10):979-987.

[29] Prasajak P. Leeanansaksiri W. Developing a new two-step protocol to generate functional hepatocytes from wharton's jelly-derived mesenchymal stem cells under hypoxic condition. Stem Cells Int. 2013;2013:762196.