共培养体系对防止体外受精小鼠胚胎因 pH 值波动而发育能力下降的作用。
The role of co-culture systems on developmental competence of preimplantation mouse embryos against pH fluctuations.
机构信息
Department of Anatomy, Afzalipour School of Medicine, Kerman University of Medical Sciences, 22 Bahman BLVD., Kerman, Iran.
出版信息
J Assist Reprod Genet. 2009 Nov-Dec;26(11-12):597-604. doi: 10.1007/s10815-009-9363-1. Epub 2009 Nov 25.
PURPOSE
To determine the effect of pH fluctuations of culture media, and the role of co-culture systems on embryo development.
METHODS
Mouse embryos were incubated in phosphate buffered solutions (PBSs) with different pH for various lengths of time. After 3 h incubation of embryos at various pH, the embryos were transferred into four media with human (HEF) and mouse (MEF) embryonic fibroblast cells, and without feeder cells; HTF and MEM-alpha. Developmental rate at day three (morula), four (expanded blastocyst) and five (hatching or hatched blastocyst) was evaluated.
RESULTS
Developmental rate at day three, four and five decreased when the incubation time at pH 6.2 and 8 increased to 3 h and more. In addition, significantly less embryos incubated at pH 6.2 and 8 developed to hatching and hatched blastocysts compared with pH 7.35. Embryos incubated at pH 6.2, co-cultured with MEF or HEF showed a significant improvement (P < 0.05) at day three in HEF compared to HTF, and at day five in MEF compared to HTF. At pH 8, a significant improvement (P < 0.05) was observed at day five in HEF and MEF compared to MEM-alpha.
CONCLUSIONS
Mouse 2-cell embryos could tolerate minor pH fluctuations, but that major pH changes affect subsequent development. Besides, feeder cells could improve embryo development, especially when embryos are prone to rise or fall in pH.
目的
确定培养基 pH 值波动以及共培养系统对胚胎发育的影响。
方法
将小鼠胚胎在不同 pH 值的磷酸盐缓冲溶液（PBS）中孵育不同时间。在不同 pH 值下孵育胚胎 3 小时后，将胚胎转移到含有人（HEF）和小鼠（MEF）胚胎成纤维细胞以及无饲养细胞的四种培养基中，即 HTF 和 MEM-alpha。评估第 3 天（桑葚胚）、第 4 天（扩张囊胚）和第 5 天（孵化或孵化囊胚）的发育率。
结果
当孵育时间延长至 3 小时及以上时，pH 值为 6.2 和 8 时，第 3、4 和 5 天的发育率下降。此外，与 pH 值 7.35 相比，在 pH 值为 6.2 和 8 下孵育的胚胎发育至孵化和孵化囊胚的数量明显减少。在 pH 值为 6.2 下孵育的胚胎与 MEF 或 HEF 共培养时，在第 3 天与 HTF 相比，HEF 中胚胎的发育有显著改善（P < 0.05），在第 5 天与 HTF 相比，MEF 中胚胎的发育有显著改善（P < 0.05）。在 pH 值为 8 时，与 MEM-alpha 相比，在第 5 天，HEF 和 MEF 中胚胎的发育有显著改善（P < 0.05）。
结论
小鼠 2 细胞胚胎可以耐受较小的 pH 值波动，但较大的 pH 值变化会影响后续的发育。此外，饲养细胞可以改善胚胎的发育，特别是在胚胎 pH 值容易上升或下降时。
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