Modified whole bone marrow adherent method for the isolation and purification of rats bone marrow mesenchymal stem cells
10.3877/cma.j.issn.2095-3232.2014.06.013
- VernacularTitle:改良全骨髓贴壁法分离与纯化大鼠骨髓间充质干细胞
- Author:
Chong SUN
1
;
Dongliang YIN
;
Huanbing ZHU
;
Kun LI
;
Jian ZHANG
Author Information
1. 中山大学附属第三医院肝移植中心
- Keywords:
Mesenchymal stem cells;
Whole bone marrow adherent method;
Rats
- From:
Chinese Journal of Hepatic Surgery（Electronic Edition）
2014;(6):56-60
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore an effective method for the isolation and purification of rats bone marrow mesenchymal stem cells (BMSCs) and to study the basic biological characteristics of BMSCs. MethodsOne healthy Sprague-Dawley (SD) rat of specific pathogen free (SPF) grade aged 4 weeks was selected, and the BMSCs were isolated using the modiifed whole bone marrow adherent method. The morphologic variation and proliferation features of BMSCs were observed under inverted phase contrast microscope, and the surface markers of BMSCs of P3 generation were detected by flow cytometry. Cell growth curves were drawn using the cell counting kit (CCK)-8 assay.ResultsTypical adherent growth was observed in the primary BMSCs with the forms of fusiform or polygon and appearance of circinate cell colony. The cell colonies fused with each other up to 80% in about 8 d. The puriifed passage cells still kept a homogeneous morphology and a high potential of proliferation. The growth curve of BMSCs indicated an S shape. The surface markers of BMSCs of P3 generation cluster of differentiation (CD) 29 and CD90 were observed positive, but CD11b and CD45 negative. ConclusionsBMSCs with high-purity, active proliferation and stable biological characteristics can be cultured using modiifed whole bone marrow adherent method. The growth curve of BMSCs presents in an S shape.
