Cryopreservation of bone marrow mesenchymal stem cells from Sprague-Dawley rats at -80 ℃

doi:10.3969/j.issn.2095-4344.2016.10.009

Abstract

Abstract:

BACKGROUND: We attempt to explore a low-cost, simple and effective way to cryopreserve bone marrow mesenchymal stem cells at -80 ℃.
OBJECTIVE: To screen the optimal cryopreservation fluid for bone marrow mesenchymal stem cells and to verify the biological features of bone marrow mesenchymal stem cells after long-term cryopreservation.
METHODS: Bone marrow mesenchymal stem cells were cultured using adherent method and the biological features and purity of cells were detected using immunofluorescence method. Bone marrow mesenchymal stem cells 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 cells. After 1, 3, 6 months of cryopreservation, the cells were resuscitated, cultured and passaged. Passage cells were identified immunofluorescence method to determine the biological features of bone marrow mesenchymal stem cells 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 cells were able to proliferate in vitro, and passage normally, indicating the cryopreserved bone marrow mesenchymal stem cells still maintain the original biological activity.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Chen Kai, Li Xin-ran, Xuan Yan. Cryopreservation of bone marrow mesenchymal stem cells from Sprague-Dawley rats at -80 ℃[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(10): 1433-1438.

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