Derivation of mesenchymal stem cell lines from menstrual blood

doi:10.3969/j.issn.2095-4344.2015.50.021

Abstract

Abstract:

BACKGROUND: Menstrual blood-derived mesenchymal stem cells are proved to be a new type of adult stem cells. To date, little is reported on the methods of establishing this kind of cell lines systematically.
OBJECTIVE: To isolate, culture, identify and finally establish menstrual blood-derived mesenchymal stem cell lines.
METHODS: Menstrual blood-derived mesenchymal stem cells were isolated from the menstrual blood of healthy female donors and cultured accordingly using density gradient centrifugation method. Cell morphology and proliferation characteristics were observed, and proliferative ability was detected by cell counting kit-8 assay. Cells were collected periodically for karyotype analysis. Expressions of cell surface antigens such as CD34, CD38, CD44, CD45, CD73, CD90, CD105 and Oct-4 were analyzed using flow cytometry. Cell pluripotency were identified by inducing adipogenesis and osteogenesis in vitro.
RESULTS AND CONCLUSION: We successfully established four menstrual blood-derived mesenchymal stem cell lines from the menstrual blood of six healthy volunteers. The cultured cell lines had a typical spindle shape and kept normal 46, XX karyotype. The cells grew rapidly and entered into the logarithmic growth phase at 48 hours after passaging. The results of flow cytometry showed that menstrual blood-derived mesenchymal stem cells were positive for CD44, CD73, CD90, CD105 and Oct-4, while negative for CD38, CD34 and CD45. Menstrual blood-derived mesenchymal stem cells were positive for oil red O staining at 22 days after adipogenic
induction and also positive for alizarin red staining at 2 weeks after osteogenetic induction. In conclusion, our study suggests that menstrual blood-derived mesenchymal stem cells with stable karyotype and pluripotency can proliferate quickly, and we can provide a new resource for stem cell therapy and regenerative medicine through establishing the menstrual blood-derived mesenchymal stem cell lines.

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

CLC Number:

R394.2

Tan Ji-chun, Li Ya-xuan, Wang Qiu-shi, Li Xiao-ni. Derivation of mesenchymal stem cell lines from menstrual blood[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(50): 8155-8160.

Figures/Tables 6

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