利用抗氧化剂促进羊水细胞培养效果的研究

目的通过添加抗氧化剂促进羊水细胞的培养效果。方法分离传代的成纤维型羊水细胞,以生长指数为检测指标,在H配方的基础上利用部分析因设计法和最陡爬坡法对促进羊水细胞生长的抗氧化剂进行筛选和优化。优化后的培养基通过原代羊水细胞培养进行效果验证。结果添加的维生素E、L-抗坏血酸、α硫辛酸和还原型谷胱甘肽等抗氧化剂皆能促进羊水细胞的生长,且具有协同效应。与H培养基相比,经最陡爬坡法优化的培养基培养的羊水细胞在克隆总数、生长指数等指标都提高3倍以上,分裂相细胞总数增长4倍以上。结论改良后的羊水细胞培养基培养效果要显著优于H配方培养基,在产前诊断应用中具有推广价值。

The study of improving the culture results of amniotic fluid cells by adding antioxidants

Objective To improve the culture results of amniotic fluid by adding antioxidants.Methods Using Fractional factorial designs combined with the steepest ascent path approach,the concentration of antioxidants added in H medium was optimized to increase the growth speed of the passage fibroblast cells from amniotic fluid.And the performance of the optimized culture medium was tested with the growth index,the numbers of clones and metaphase of primary amniotic fluid cells culture.Results Vitamin E,L-ascorbic acid,α-lipoic acid and reduced GSH can all promote the growth of the amniotic fluid cells with synergic effect.Total number of clones,growth index,and total number of metaphase cells from amniotic fluid cells cultured in medium optimized by the steepest ascent path approach are all 3 times higher than those from H medium.Conclusion The optimization method of adding antioxidants in H medium can improve significantly the results of amniotic fluid cells culture and the optimized culture medium is applicable to the prenatal diagnosis.