ENVIRONMENTAL VARIABLES INFLUENCING INVITRO DEVELOPMENT OF HAMSTER 2-CELL EMBRYOS TO THE BLASTOCYST STAGE
ENVIRONMENTAL VARIABLES INFLUENCING INVITRO DEVELOPMENT OF HAMSTER 2-CELL EMBRYOS TO THE BLASTOCYST STAGE
复制标题
DOI：
10.1095/biolreprod43.3.404
复制
发表时间：
1990-09-01
影响因子：
3.6
通讯作者：
BAVISTER, BD
中科院分区：
文献类型：
--
作者：
MCKIERNAN, SH;BAVISTER, BD
本研究系统分析了影响2细胞仓鼠胚胎体外发育至囊胚期的环境变量。实验使用化学定义的（无蛋白）培养基（HECM-2）进行。检测的物理化学变量包括温度、CO2、HCO3-、Ca2+、Mg2+、K+和O2的浓度、硅油覆盖层的存在以及渗透压。在硝基中发育的最佳温度为37.5%℃；较低的温度有抑制作用。在测试范围内，NaHCO3、CaCl2、MgCl2和KCl浓度的变化，或Ca2+:Mg2+和Na+:K+的比例的变化，对囊胚发育没有显著影响。提高CO2浓度（7.5%和10%），降低O2浓度（10%和5%），以及在培养基上覆盖硅油，都能显著促进囊胚期的发育。在没有油覆盖层的情况下，囊胚发育的减少不是由渗透压的增加引起的。250 ~ 350 mOsmols的渗透压对卵裂期胚胎没有影响，但在0.5 mOsmols时囊胚发育受到抑制。325 mOsmols。在优化条件（37.5℃，10% CO2, 25 mM HCO3-, 2.0 mM Ca2+, 0.5 mM Mg2+, 3.0 mM K+, 10% O2, 250-300 mOsmols，硅油覆盖）下，51-57%的2细胞仓鼠胚胎发育到囊胚期。这比以前的“标准”培养条件有了显著的改善，标准培养条件支持26%的2细胞仓鼠胚胎囊胚发育。这里描述的培养系统提供了足够的基线反应，以支持对仓鼠胚胎发育调节的详细调查。
This study is a systematic analysis of environmental variables influencing development of 2-cell hamster embryos to the blastocyst stage in vitro. Experiments were done using a chemically defined (protein-free) culture medium (HECM-2). Physicochemical variables examined were temperature, the concentrations of CO2, HCO3-, Ca2+, Mg2+, K+, and O2, the presence of a silicone oil overlay, and osmotic pressure. The optimal temperature for development in nitro was 37.5%C; lower temperatures were inhibitory. There was no significant effect on blastocyst development of alterations in the concentrations of NaHCO3, CaCl2, MgCl2, and KCl, or in the ratio of Ca2+ :Mg2+ and Na+ :K+, over the ranges tested. Development to the blastocyst stage was significantly stimulated by increased CO2 concentrations (7.5% and 10%), by reduced O2 concentrations (10% and 5%), and by the presence of silicone oil overlying the culture medium. Reduction of blastocyst development in the absence of an oil overlay was not caused by increased osmotic pressure. Cleavage stage embryos were not affected by osmolalities ranging from 250 to 350 mOsmols, but blastocyst development was inhibited at .gtoreq.325 mOsmols. Under optimized conditions (37.5.degree.C, 10% CO2, 25 mM HCO3-, 2.0 mM Ca2+, 0.5 mM Mg2+, 3.0 mM K+, 10% O2, 250-300 mOsmols, with silicone oil overlay), 51-57% of 2-cell hamster embryos developed to the blastocyst stage. This represents a significant improvement over previous "standard" culture conditions, which supported development of 26% blastocysts from 2-cell hamster embryos. The culture system described here provides an adequate baseline response to support detailed investigations into the regulation of embryo development in the hamster.