Derivation of mesenchymal stem cell lines from menstrual blood / 中国组织工程研究

ABSTRACT

BACKGROUND:Menstrual blood-derived mesenchymal stem cels are proved to be a new type of adult stem cels. To date, little is reported on the methods of establishing this kind of cel lines systematicaly. OBJECTIVE:To isolate, culture, identify and finaly establish menstrual blood-derived mesenchymal stem cel lines. METHODS:Menstrual blood-derived mesenchymal stem cels were isolated from the menstrual blood of healthy female donors and cultured accordingly using density gradient centrifugation method. Cel morphology and proliferation characteristics were observed, and proliferative ability was detected by cel counting kit-8 assay. Cels were colected periodicaly for karyotype analysis. Expressions of cel surface antigens such as CD34, CD38, CD44, CD45, CD73, CD90, CD105 and Oct-4 were analyzed using flow cytometry. Cel pluripotency were identified by inducing adipogenesis and osteogenesis in vitro. RESULTS AND CONCLUSION:We successfuly established four menstrual blood-derived mesenchymal stem cel lines from the menstrual blood of six healthy volunteers. The cultured cel lines had a typical spindle shape and kept normal 46, XX karyotype. The cels 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 cels were positive for CD44, CD73, CD90, CD105 and Oct-4, while negative for CD38, CD34 and CD45. Menstrual blood-derived mesenchymal stem cels 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 cels with stable karyotype and pluripotency can proliferate quickly, and we can provide a new resource for stem cel therapy and regenerative medicine through establishing the menstrual blood-derived mesenchymal stem cel lines.