冷冻预处理温度对胎兔许旺细胞免疫原性的影响

目的探讨不同维持温度冷冻预处理超深低温保存对胎兔许旺细胞免疫原性的影响。方法设未冷冻处理细胞为对照组,实验组细胞设计冷冻维持温度为-30℃,-35℃,-40℃,-45℃,-50℃,-55℃,-60℃,-65℃,-70℃,10%二甲基亚砜（DMSO）为低温保护剂,对胎兔许旺细胞进行冷冻预处理,液氮中保存48h,复温,培养48h后培养液中加入IFN-γ（100U/ml）继续培养72h,流式细胞仪检测许旺细胞MHC-Ⅱ分子的表达率。结果流式细胞仪检测显示各实验组-30℃至-70℃许旺细胞MHC-Ⅱ分子的表达率依次分别是：7.59±0.91,10.43±0.90,14.35±0.73,16.80±0.73,15.41±0.71,14.50±0.73,10.92±0.89,8.36±0.68和5.26±0.76,对照组为35.98±1.06,统计学显示实验组MHC-Ⅱ表达率和对照组之间有显著性差异（P〈0.01）。结论不同维持温度冷冻预处理超深低温保存能降低胎兔许旺细胞的免疫原性。

Effect of pre-treatment freezing temperature before cryopreservation on the immunogenicity of embryonic rabbit's Schwann cells.

Objective To investigate the effect of pre-treatment freezing temperature before cryopreservation on the immunogenicity of embryonic rabbit＇s Schwann cells.Methods Schwann cells without cryopreservation were set as blank control.Schwann cells in experimental groups were frozen at-30℃,-35℃,-40℃,-45℃,-50℃,-55℃,-60℃,-65℃ and-70℃ respectively and treated with a cryoprotectant at the same time such as 10% dimethyl sulfoxide（DMSO）,preserved in liquid nitrogen for 48 h,cultured for 48 h after rapid rewarming,followed by culture with IFN-γ（100 U/mL） for 72 h.The expression of MHC-Ⅱ in Schwann cells was detected by flow cytometry.Results The expression of MHC-Ⅱ in blank control was 35.98±1.06,and that in Schwann cells pre-treated with different freezing temperature from-30℃ to-70℃ at a-5℃ scale was 7.59±0.91,10.43±0.90,14.35±0.73,16.80±0.73,15.41±0.71,14.50±0.73,10.92±0.89,8.36±0.68 and 5.26±0.76 respectively.The expression of MHC-Ⅱ in experimental groups was significantly lower than in blank control group（P0.01）.Conclusion Pre-treatment freezing temperature before cryopreservation can decrease the immunogenicity of embryonic rabbit＇s Schwann cells.