Effect of different polyols on the cryopreservation of mouse embryonic stem cells
- VernacularTitle:不同多羟基化合物对小鼠胚胎干细胞低温贮存的影响
- Author:
Dandan YAO
;
Chunhui YUAN
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;11(3):576-578
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: The long-term cryopreservation of embryonic stem cells is an important link in its researches and application. Traditional protectants of cryopreservation often contain dimethyl sulfoxide, glycerine, animal serum, etc., but they cannot effectively ensure the survival rate of some cells, and even damage some functions of the cells.OBJECTIVE: To develop the suitable protectant for the cropreservation of mouse embryonic stem cells. DESIGN: An observational study.SETTINGS: Department of Physiology, Conghua College of Guangzhou Medical College; Sinocells Bio Technologies Co.,Ltd.MATERIALS: The experiment was carried out in the laboratory of Sinocells Bio Technologies Co.,Ltd. from October 2004 to September 2005. Mouse cell line mES-1 was given as a present by the Research Room of Cell Biology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences of Chinese PLA.METHODS: The cells collected from cultured mES-1, a mouse embryonic stem cell line. Dimethyl sulfoxide was used as the main protectant for cryopreservation. There were four different groups according to different protectants. In the control group, the protectant for cryopreservation contained DMEM (high sugar), 10% dimethyl sulfoxide, 10% FBS,10 mg/mL ascorbicacid, 0.18 mg/mL inositol and 0.44 mg/mL folic acid; Besides, 7.56 mg/mL mannose in the mannose group, 34.23 mg/mL mycose in the mycose and 41.94 mg/mL saccharu in the saccharu group were supplemented respectively. The effects of different polyols on the survival rate and ability of multiple differentiation of mouse embryonic stem cells after cryopreservation were observed.MATN OUTCOME MEASURES: Formation rate of mouse embryonic stem cells colony; Formation rate of embryoid bodies.were all significantly decreased as compared with that before cryopreservation [control group: (24.0±8.8)%; mannose group: (42.0±10.1)%; mycose group: (84.0±8.2)%; saccharu group (70.0±14.2)%; before cryopreservation: (95.0±4.7)%,embryoid bodies in the mycose group was higher than those in the control group and mannose group [(90.0±5.2)%,(80.0±6.9)%, (82.0±9.6)%, P＜0.05], but had no significant difference as compared with that in the saccharu group.CONCLUSION: The protectant for cryopreservation by taking mycose as the main component can effectively maintain the abilities of self-renewing and multiple differentiation of mouse embryonic stem cells.
