Primary culture of murine spleen-derived mesenchymal stem cells by explant culture
10.3969.j.issn.1671-7856.2016.10.011
- VernacularTitle:组织块法分离培养小鼠脾脏间充质干细胞
- Author:
Li DING
;
Heng ZHU
;
Haihong ZHANG
;
Yang YANG
;
Dongmei HAN
;
Zhidong WANG
;
Xiaoli ZHENG
;
Lei DONG
;
Hongmin YAN
;
Jing LIU
;
Ling ZHU
;
Mei XUE
;
Zikuan GUO
;
Hengxiang WANG
- Publication Type:Journal Article
- Keywords:
Tissue explant;
Cell culture;
Spleen;
Mesenchymal stem cells;
Osteoblast differentiation;
Adipocyte differentiation;
Mice
- From:
Chinese Journal of Comparative Medicine
2016;26(10):56-60
- CountryChina
- Language:Chinese
-
Abstract:
Objective This study aimed to establish a reliable primary culture protocol for preparing murine spleen-derived mesenchymal stem cells ( MSCs) by tissue explant culture.Methods Healthy mouse spleens were crushed by syringe handle to harvest spleen mesenchymal tissues.Then the tiny pieces of spleen tissue were digested by collagenase II before seeded into culture flasks.The morphological characteristics of spleen tissue-derived cells were observed under the inverted microscope.Further, the surface antigen profile of the cells was analyzed by flow cytometry (FACS).The cells were induced to differentiate into osteoblasts and adipocytes.Results The murine spleen-derived MSCs exhibited a spindle-shaped appearance.The FACS results showed that the spleen-derived MSCs highly expressed CD29, CD44, CD105 and Sca-1, but weakly expressed CD11b, CD34, CD45 and Ia. In addition, the spleen-derived MSCs steadily differentiated into osteoblasts and adipocytes in the induction medium.Conclusions A method of primary culture of murine spleen-derived MSCs by explant culture is successfully established.The harvested MSCs exhibit high purity and cell proliferation ability, and provide a reliable cell model for related researches.
