Cryopreservation of bone marrow mesenchymal stem cells from Sprague-Dawley rats at-80℃
10.3969/j.issn.2095-4344.2016.10.009
- VernacularTitle:SD大鼠骨髓间充质干细胞在-80℃低温条件下的冻存
- Author:
Kai CHEN
;
Xinran LI
;
Yan XUAN
- Publication Type:Journal Article
- Keywords:
Bone Marrow;
Mesenchymal Stem Cels;
Dimethyl Sulfoxide;
Cryopreservation;
Tissue Engineering
- From:
Chinese Journal of Tissue Engineering Research
2016;20(10):1433-1438
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:We attempt to explore a low-cost, simple and effective way to cryopreserve bone marrow mesenchymal stem cels at-80℃.
OBJECTIVE:To screen the optimal cryopreservation fluid for bone marrow mesenchymal stem cels and to verify the biological features of bone marrow mesenchymal stem cels after long-term cryopreservation.
METHODS: Bone marrow mesenchymal stem cels were cultured using adherent method and the biological features and purity of cels were detected using immunofluorescence method. Bone marrow mesenchymal stem cels were cryopreserved in the cryoprotectant medium containing low-sugar DMEM, fetal bovine serum and dimethyl sulfoxide at different proportions at-80℃ for a short term. Then, the optimal cryoprotectant was selected to storage the bone marrow mesenchymal stem cels. After 1, 3, 6 months of cryopreservation, the cels were resuscitated, cultured and passaged. Passage cels were identified immunofluorescence method to determine the biological features of bone marrow mesenchymal stem cels cryopreserved at-80℃.
RESULTS AND CONCLUSION:Cryoprotectant medium of 80% DMEM+10% fetal bovine serum+10% dimethyl sulfoxide was suitable for cryopreserving MSCs at -80℃, and resuscitated cels were able to proliferate in vitro, and passage normaly, indicating the cryopreserved bone marrow mesenchymal stem cels stil maintain the original biological activity.
