Isolation and identification of nucleus pulposus mesenchymal stem cells from tissues removed by percutaneous endoscopic lumbar discectomy
10.3969/j.issn.1000-4718.2017.06.032
- VernacularTitle:从脊柱内镜手术摘除组织中分离髓核间充质干细胞及其生物学特征鉴定
- Author:
Yupan SHANG
;
Hao WU
;
Xiaoli ZENG
;
Lekhang CHEANG
;
Jun YU
;
Qianlong XIAO
;
Mei TU
;
Jiaqing ZHANG
- Keywords:
Percutaneous endoscopic lumbar discectomy;
Explant culture method;
Human nucleus pulposus mesenchymal stem cells
- From:
Chinese Journal of Pathophysiology
2017;33(6):1147-1152
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To explore a novel method to isolate human nucleus pulposus mesenchymal stem cells (hNP-MSCs) in vitro and to identify their biological characteristics.METHODS:The explant culture method was employed to isolate hNP-MSCs from nucleus pulposus tissue obtained by percutaneous endoscopic lumbar discectomy (PELD).The isolated cells were passaged for purification and cultured in vitro followed by morphological observation.The cell proliferation ability was detected by CCK-8 assay.Growth curves of the cells were drawn and surface antigens were detected by flow cytometry.The cells at the 3rd~6th passages were induced for adipogenic, osteogenic and chondrogenic differentiation, and examined by oil red O staining, alizarin red staining and Alcian blue staining.RESULTS:The cells with self-renewal were obtained from nucleus pulposus tissue obtained by PELD.The results of flow cytometry analysis revealed that the cells were positive for CD29, CD44, CD90, CD73 and CD105, but negative for CD34 and CD45.The proliferative capacity was consistent with the growth characteristics of MSCs and multilineage differentiation potential was identified.CONCLUSION:A novel method to efficiently isolate and culture hNP-MSCs,PELD combined with explant culture method,was established, which would promote the study of regenerative medicine based on hNP-MSCs.
