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.
ABSTRACT
BACKGROUND:Adipose-derived stem cels are pluripotent stem cels developed from the mesoderm, which can differentiate into different lineages induced by specific growth factors and under certain environmental conditions. OBJECTIVE: To describe the induced differentiation and identification of adipose-derived stem cels in detail. METHODS:A computer-based search of Wanfang and PubMed databases was performed for relevant articles published from 2005 to 2014 using the keywords of “adipose derived stem cels, induced, differentiation” in Chinese and English, respectively. According to inclusion and exclusion criteria, 37 articles were selected in result analysis. RESULTS AND CONCLUSION:Adipose-derived stem cels can differentiate into chondrocytes under the induction of ascorbic acid, insulin, dexamethasone, and transforming growth factor β. Recipe for adipogenic induction medium consists of 3-isobutyl-1-methylxanthine (IBMX), insulin, dexamethasone, indomethacin; and commonly used inducers for osteogenic differentiation include dexamethasone or vitamin D3, ascorbic acid,β-glycerophosphate. Basic fibroblast growth factor, epidermal growth factor and vitamin B27 may be combined to induce the differentiation of adipose-derived stem cels into neurons. 5-Azacytidine acts as a commonly used factor for inducing the differentiation of adipose-derived stem cels into cardiomyocytes. Combination of vascular endothelial growth factor and basic fibroblast growth factor can induce adipose-derived stem cels to differentiate into vascular endothelial cels. With the rapid development of molecular biology and cel biology, research on the differentiation of adipose-derived stem cels wil be more in-depth. Based on the current observation of adipose-derived stem cel differentiation, internal molecular mechanism as wel as genes and proteins for regulation of adipose-stem cel plasticity should be explored in depth.