Isolation,Culture,Identification and Induced Differentiation of Mesenchymal Stem Cells from Human Umbilical Cord
10.3969/j.issn.1671-7414.2015.03.019
- VernacularTitle:人脐带间充质干细胞分离培养鉴定及诱导分化实验
- Author:
Na XIE
;
Ming LOU
;
Guozhou RAO
;
Yong ZHU
;
Chengfang TANG
;
Lin WANG
- Publication Type:Journal Article
- Keywords:
umbilical cord;
wharton glue;
stem cells;
induced differentiation
- From:
Journal of Modern Laboratory Medicine
2015;(3):67-70
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the method of isolation,culture and identify biological characterization of mesenchymal stem cells from human umbilical cord (hUCMSCs);and study their multiple differentiation potency.Methods Stem cells from human umbilical cord were cultured by enzyme Wharton jelly method in vitro.The surface markers were identified by flow cytometry.Multi-differentiation capacity was identified by osteogenic and adipogenic differentiation.ALP was detected with Calcium cobalt staining.The mineralized ability in vitro was measured with Alizarin red staining.Theadipocyte differen-tiation ability was measured with oil red-O staining.Results Flow cytometry analysis revealed that CD73 (92.45%),CD90 (95.45%)and CD105 (96.45%)were highly expressed on these cells’surface,while CD34 (1.07%)were negative ex-pressed.Cells were cultured with induced-osteogenic medium after 3 weeks,ALP staining in the cytoplasm of black parti-cles,and a large amount of mineralized nodules within cells was observed after 4 weeks.Cells were cultured with induced-adi-pogenic medium after 2 weeks,the majority of these cells were round,oil red O staining of lipid droplets generated within cells was observed.Conclusion Mesenchymal stem cells from human umbilical cord have the potential of multi-directional differentiation.These cells could be induced to differentiate into adipocytes and osteoblasts,which laid the foundation for clinical stem cell therapy research source of seed cells.
