卵胞浆内注射前圆形精子细胞核分离及保存培养基的比较
Comparison of the media for isolation and storage of round spermatid nuclei before intracytoplasmic injection.
作者信息
Suzuki K, Yanagida K, Yanagimachi R
机构信息
Department of Anatomy and Reproductive Biology, University of Hawaii School of Medicine, Honolulu 96822, USA.
出版信息
J Assist Reprod Genet. 1998 Mar;15(3):154-7. doi: 10.1023/a:1023013022295.
PURPOSE
This study investigated whether K(+)-rich medium is better than pure NaCl solution or Na(+)-rich cell culture medium for handling round spermatid nuclei prior to injection into oocytes (ROSNI).
METHODS
Round spermatids of the mouse were isolated and stored in isotonic NaCl, a cell culture medium (CZB), or a nucleus isolation medium (NIM) before injection into oocytes. The rates of normal fertilization, embryonic development in vitro, and birth of normal offspring after transfer of embryos to foster mothers were determined.
RESULTS
In vitro development of ROSNI-produced zygotes to blastocysts was the same when "naked" spermatid nuclei were exposed briefly to three media. However, a long (60-min) exposure of the nuclei to Na(+)-rich medium was detrimental. In K(+)-rich NIM "naked" spermatid nuclei best retained their ability to participate in normal embryonic development.
CONCLUSION
NIM was better than Na(+)-rich medium for retaining isolated spermatids competent to participate in normal embryonic development.
目的
本研究调查了在将圆形精子细胞核注入卵母细胞（ROSNI）之前，富含钾离子的培养基是否比纯氯化钠溶液或富含钠离子的细胞培养基更适合处理圆形精子细胞核。
方法
从小鼠中分离出圆形精子细胞，并在注入卵母细胞之前，将其保存在等渗氯化钠、细胞培养基（CZB）或细胞核分离培养基（NIM）中。测定正常受精率、体外胚胎发育率以及将胚胎移植到代孕母鼠后正常后代的出生率。
结果
当“裸露”的精子细胞核短暂暴露于三种培养基时，ROSNI产生的合子体外发育至囊胚的情况相同。然而，细胞核长时间（60分钟）暴露于富含钠离子的培养基是有害的。在富含钾离子的NIM中，“裸露”的精子细胞核最能保留其参与正常胚胎发育的能力。
结论
在保留有能力参与正常胚胎发育的分离精子细胞方面，NIM比富含钠离子的培养基更好。
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