摘要
将胎儿成纤维细胞悬浮于不同的冷冻保护液中 ,在不同的温度下进行冷冻保存试验。结果显示 ,当保护液中含有 90 0 m L / L胎牛血清 (FBS)时 ,- 196℃下保存的山羊胎儿成纤维细胞解冻后的细胞贴壁率高于- 80℃ ,且前者贴壁细胞增殖也较后者快 ,二者间差异显著 ;而在 - 2 0℃条件下只有一小部分细胞能够存活 ,与前两者相比差异极显著 ,说明 - 2 0℃不适合冷冻胎儿细胞。冷冻保护液中含有 10 0 m L / L二甲基亚砜 (DMSO)的保护效果明显好于 5 0 m L / L DMSO,二者间差异显著。用组织块法培养的细胞 ,在两种培养基 DMEM和 M199中添加 10 0 m L / L FBS和 2 mm ol/ L 2 -巯基乙醇 (2 - Me) ,其组织块成活率及细胞生长速度均高于单纯添加 10 0 m L / LFBS,同时证明 DMEM和 M199都能用于胎儿成纤维细胞的体外培养。以 DMEM培养液为基础 ,分别添加 2mm ol/ L 2 - Me,5 m L / L FBS,10 0 m L / L FBS和 2 m mol/ L 2 - Me+10 0 m L / L FBS对山羊胎儿成纤维细胞进行体外培养 ,结果显示 ,细胞在 10 0 m L / L FBS和 2 mm ol/ L 2 - Me+10 0 m L / L FBS培养液中生长迅速 ,两者间无明显差异 ,2 - Me的存在促进了细胞的增殖 ;两者都与添加 2 m mol/ L 2 - Me组存在显著差异 ;添加 2 m mol/ L 2 - Me和 5m L / L
Goat fetal fibroblasts were frozen at different tem perature with various protecting solution in order to find a proper procedure to preserve its rare resources for further study both in transgenic and cloning practice. The result showed that fetal fibroblasts froze at- 196℃ with 90 0 m L/ L fetal bovine serum(FBS) grew faster than that at- 80℃ after thawing(P<0 .0 5 ) .- 2 0℃ was not an effective temperature in frozen cells.10 0 m L/ L dimethyl sulfoxide(DMSO) contained in frozen solution was better than 5 0 m L/ L in im proving protection effec- tiveness(P<0 .0 5 ) .In vitro culture of fetal fibroblasts experiment showed that both DMEM and M199can sup- port cell growth and proliferation and that a supplem entation of 2 mm ol/ L 2 - m ercaptoethanol (2 - Me) was better than not in improving tissue vitality and growth speed.In cell culture procedure,2 - Me could improve the effect of 10 0 m L / L FBS in stim ulating cell proliferation.And individual 2 - Me supplementation in the culture medium could stimulate cell growth.As a result,2 - Me was more likely a cell proliferation stim ulator than an oxidation retarding factor.High serum concentration10 0 m L/ L was vital for cell proliferation,serum starvation(5 m L/ L FBS) could not only inhibit cell division but also cause cell loss during long term culture.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2004年第4期55-58,共4页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家 8 63计划项目"家畜体细胞克隆技术体系的建立和应用"( 2 0 0 1AA2 13 0 81)
