Optimization of the method for isolating and culturing rat mesenchymal stem cells.
- Author:
Xulong ZHU
1
,
2
;
Tan YAN
;
Weijie YAO
;
Yongheng WANG
;
Chong CHENG
;
Junxi XIANG
;
Yi LV
;
Qingdong GAO
;
Jianhui LI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Cell Separation; Flow Cytometry; Mesenchymal Stromal Cells; cytology; Rats
- From: Journal of Southern Medical University 2014;34(11):1621-1631
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo optimize the protocols for isolation and culture of mesenchymal stem cells from rat bone marrow (BMSCs).
METHODSBMSCs were isolated by adherence to plastic with frequent medium change and reduced trypsinization time. The cell growth curves were drawn and the surface markers of BMSCs were detected by flow cytometry. The cells were induced to differentiate into osteogenic, adipogenic, hepatic and cholic lineages.
RESULTSThe cells isolated using this method were positive for CD29, CD44, and CD90 and negative for the hematopoietic surface markers CD45. The osteogenic and adipogenic differentiation of the BMSCs was verified by alkaline phosphatase staining, Alizarin red staining and Oil red staining. The cell subcultures up to passage 10 maintained capacities of differentiation into osteogenic and adipogenic lineages. The BMSCs induced with sequential addition of growth factors, cytokines and hormones differentiated into cells expressing hepatocyte- and cholangiocyte-specific markers.
CONCLUSIONThe optimized method allows efficient isolation of homogenous populations of MSCs from rat bone marrow, which can be induced into multiple cell lineages.
