Isolation and cultivation of human placental chorionic-derived mesenchymal stem cells: optimization of the tissue explants method
10.3969/j.issn.2095-4344.2017.21.018
- VernacularTitle:人胎盘绒毛膜间充质干细胞分离培养:组织块法的优化
- Author:
Yulin JIN
;
Jieying WU
;
Yan LU
;
Jinsong CHEN
;
Fatao LI
;
Jie TANG
;
Dong LIU
;
Qihua LIANG
;
Yan LI
;
Xuewei TANG
;
Guie XIE
;
Shaoqing WU
- From:
Chinese Journal of Tissue Engineering Research
2017;21(21):3388-3393
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:There are a lot of studies on isolation and culture methods of human placental chorionic-derived mesenchymal stem cells (hpcMSCs), but how to simply and efficiently harvest a large amount of primary MSCs has not been resolved. OBJECTIVE:To optimize the tissue explants method of isolating and culturing hpcMSCsin vitro. METHODS:Human placental chorionic villi were collected from full-term deliveries under aseptic condition and isolated by electric homogenizer. hpcMSCs were prepared by tissue explants method. The fluid and tissue of the primary culture flask and douching normal saline of the initial culture were centrifuged and prepared for secondary culture. RESULTS AND CONCLUSION: It saved time and effort to treat human placental chorionic villi with electric homogenizer, with good effects on tissue dispersion and removal of red blood cells. The average time of cell acquisition in initial culture and secondary culture was (17.73±1.14) and (10.03±1.30) days, respectively. The yields of primary cultured cells in initial culture and secondary culture were (6.97±0.98)×105 and (13.82±1.44)×105per Φ100 mm culture dish, respectively. The adherent cells showed fibroblast-cell-like shape, which were in parallel or circinate arrangement. Highly expressed CD73, CD105 and CD90 could be detected in the third generation of hpcMSCs, but CD34, CD45, CD14, CD19 and HLA-DR were negative. Following induction, alizarin red staining and oil red O staining produced a strong reaction in cells. In a word, the optimized method is a simple and efficient method for obtaining a large amount of primary hpcMSCs.
