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基质胶能否替代Vero细胞促进小鼠胚胎的体外发育?

Can Matrigel substitute for Vero cells in promoting the in-vitro development of mouse embryos?

作者信息

Carnegie J, Claman P, Lawrence C, Cabaca O

机构信息

GOAL Programme, University of Ottawa, Ontario, Canada.

出版信息

Hum Reprod. 1995 Mar;10(3):636-41. doi: 10.1093/oxfordjournals.humrep.a136002.

Abstract

The influences of Vero cells and the basement membrane substratum for these cells (Matrigel) on the rate of hatched blastocyst formation from mouse zygotes in vitro were compared. Zygotes obtained from C57BL/6 x BALB/c F1 females pretreated with pregnant mare's serum gonadotrophin/human chorionic gonadotrophin mated with BDF1 males were cultured (120 h) in human tubal fluid medium supplemented 0.5% with bovine serum albumin. The rates of early hatching and hatched blastocyst formation at 96 and 120 h of culture were expressed as the percentage of 2-cell embryos visualized after the initial 24 h. The rate of total blastocyst formation did not differ between treatment groups. However, < 10% of embryos cultured for 96 h in medium alone advanced to the hatching stage compared with 35-40% of blastocysts cultured with Vero cells or with Matrigel alone. Similarly, by 120 h of culture, only 20% of embryos cultured in medium alone developed to hatching or hatched blastocysts compared with > 70% for those embryos co-cultured with Vero cells or with Matrigel. In conclusion, Vero cells improved the rate of development of mouse embryos to hatched blastocysts during serum-free culture. Similar improvements were seen in the presence of Matrigel alone; Matrigel is the basement membrane substratum used for the Vero cells. Further studies on the means whereby Matrigel promotes early embryonic development (e.g. appropriate combination of basement membrane-associated growth factors) may lead to a safe, defined medium preparation for the stimulation of in-vitro development of human embryos.

摘要

比较了Vero细胞及其基底膜基质(基质胶)对小鼠受精卵体外孵化囊胚形成率的影响。从用孕马血清促性腺激素/人绒毛膜促性腺激素预处理的C57BL/6×BALB/c F1雌性小鼠获得的受精卵,与BDF1雄性小鼠交配后,在补充有0.5%牛血清白蛋白的人输卵管液培养基中培养(120小时)。培养96小时和120小时时的早期孵化率和孵化囊胚形成率表示为最初24小时后可见的2细胞胚胎的百分比。各处理组之间的总囊胚形成率没有差异。然而,单独在培养基中培养96小时的胚胎中,只有不到10%进入孵化阶段,而与Vero细胞或单独与基质胶共培养的囊胚中这一比例为35%-40%。同样,到培养120小时时,单独在培养基中培养的胚胎中只有20%发育到孵化或孵化囊胚阶段,而与Vero细胞或基质胶共培养的胚胎中这一比例超过70%。总之,在无血清培养期间,Vero细胞提高了小鼠胚胎发育到孵化囊胚的速率。单独存在基质胶时也观察到类似的改善;基质胶是用于Vero细胞的基底膜基质。对基质胶促进早期胚胎发育的方式(例如基底膜相关生长因子的适当组合)的进一步研究,可能会导致一种安全、明确的培养基制剂,用于刺激人类胚胎的体外发育。

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