不同样品保存液和保存条件对动物细胞体外培养的影响

本研究取矮脚油鸡7日龄胚胎作为试验材料，用MEM、全培养基和PBS 3种不同的液体对鸡胚组织进行保存，同时在不同保存温度条件下按不同保存时间梯度，采用组织块贴壁培养法进行体外细胞培养。对细胞的保存液变化，细胞形态、细胞生长等情况进行研究，分析样品保存条件对细胞体外培养和细胞系构建的影响。结果发现，4 ℃保存的组织块培养效果明显优于室温保存；无论在4 ℃或常温，基础培养基保存的组织块培养效果明显优于保存液，全培养基效果最好。常温保存48 h以内3种保存液培养效果差别甚微，48 h后培养效果均急剧下降，96 h后保存在PBS中的组织失活，未长出细胞。本研究结果为建立细胞系采集样品的保存提供了一定的参考。

The Effect of Various Sample Conservation Medium and Condition on Cell Culture in Vitro

Seven-days-old embryo of Short-feet Chicken was studied in present research, and the tissues of chicken embryo were conserved by three different medium. The variation of tissue conservation culture medium, cell morphology and cell growth were studied by tissue culture for different conservation time at different conservation temperature to analyze the effect of tissue conservation condition on cell culture in vitro. The results indicated that the cells from tissue conserved at 4 ℃ were better than conserved at room temperature; the cells from tissue preserved in basic culture medium MEM and whole culture medium were obviously better than preserved in PBS either at 4 ℃ or at room temperature, and the whole culture medium was the best conservation medium; there is no significant difference among these three conservation mediums when preserved in 48 hours at room temperature, while the culture become bad deeply when preserved after 48 hours, and the tissue died and no cell grew in PBS after 96 hours. The present study supplied some reference to the conservation of tissue and the construction of cell line.