In vitro proliferation and passage of bone marrow mesenchymal stem cells impact homing-associated factors
10.3969/j.issn.2095-4344.2013.40.013
- VernacularTitle:体外扩增传代对骨髓间充质干细胞归巢相关因子的影响
- Author:
Wen XU
;
Chen TIAN
;
Fang LI
;
Bing XIA
;
Qing GUO
;
Yizhuo ZHANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2013;(40):7102-7109
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:The homing ability of mesenchymal stem cells after transplantation can decrease along with culture and passage in vitro. And the decline of homing abilities can further influence the implantation of mesenchymal stem cells in the target tissue, thus seriously affecting the repair effect.
OBJECTIVE:To investigate the effect and its related mechanisms by which in vitro culture and passage affect the homing ability of mesenchymal stem cells.
METHODS:Mesenchymal stem cells were isolated from the bone marrow using Ficol density gradient centrifugation, and then purified using adhesion method. The mesenchymal stem cells were cultured into the seventh generations with the normal cultural condition, and the morphological features of the 3rd, 5th and 7th generations of mesenchymal stem cells were observed. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide chromatometry was used to detect the growth feature of the 3rd, 5th and 7th generations of mesenchymal stem cells, and the growth curve was drawn. Real-time PCR was used to detect the expression of CXCR4, CXCR6, CXCL12, CD44 in the 3rd, 5th and 7th generations of mesenchymal stem cells, 2-△△Ct was calculated to get the relative value of each target gene, and the differences in expression of CXCR4, CXCR6, CXCL12, CD44 between different generations were compared.
RESULTS AND CONCLUSION:Mononuclear cells could be obtained from the bone marrow by using Ficol density gradient centrifugation. High-purity mesenchymal stem cells could be obtained using adherent method. The ability of in vitro growing was strong, but fol owing the passage, the cellmorphology became wider and shorter. And the proliferation rate, the overal proliferated multiple and the expression of homing related factors decreased fol owing the passage. The expression of CXCR4, CXCR6, CXCL12 and CD44 of mesenchymal stem cells decreased fol owing the passage. The homing ability of mesenchymal stem cells was decreased fol owing the passage, and may be relevant with the lower expression of CXCR4, CXCR6, CXCL12 and CD44 in cultured mesenchymal stem cells.
