大鼠和小鼠囊胚在体外着床前及着床期间内细胞团发育的免疫外科研究。

Immunosurgical studies on inner cell mass development in rat and mouse blastocysts before and during implantation in vitro.

作者信息

Spielmann H, Jacob-Müller U, Beckord W

出版信息

J Embryol Exp Morphol. 1980 Dec;60:255-69.

Abstract

Eighty per cent of rat blastocysts (Wistar, SW72) cultured for 96 h in NCTC-109 supplemented with fetal calf serum (FCS) hatched from the zona pellucida and developed a trophoblast giant cell lyer. Thirty seven per cent from the rat blastocysts developed an inner cell mass (ICM) which, in about 7% consisted of two germ layers (ectoderm and endoderm) compared to 84% in NMRI mice. A significantly better ICM development was obtained with cultured rat blastocysts that had hatched in vivo. Similar to the in vivo situation LDH-5 was present in rat blastocysts after implantation in NCTC-109-FCS. Differentiation of C57BL mouse blastocysts in NCTC-109-FCS proceeded as poorly as in the rat. ICM development of rat and mouse blastocysts in NCTC-109-FCS was studied in detail. ICMs of the two species were isolated immunosurgically using complement from different species, e.g. human, rat and rabbit complement, since guinea-pig complement did not lyse trophectoderm cells of rat blastocysts. All immunosurgically isolated rat ICMs degenerated within 48 h, but mouse ICMs isolated with rat or rabbit complement developed significantly better than mouse ICMs isolated with guinea-pig complement. Determinations of the blastocyst total cell number (BTCN) and of the cell number of immunosurgically isolated ICMs were performed in rat and mouse blastocysts to investigate growth kinetics of the ICM before implantation in vitro. In the mouse an exponential increase in both BTCN and cell number of the ICM was observed during the 48 h before implantation in NCTC-109-FCS and also during the 16-24 h before implantation in vivo. In the rat, doubling of the BTCN was found only during the first 24 h in NCTC-109-FCS and there was hardly any increase in the cell number of the ICM during the first 48 h in culture. ICM growth of blastocysts in NCTC-109-FCS is therefore, stimulated in the mouse before and after implantation and in the rat it is inhibited already before implantation.

摘要

在补充有胎牛血清（FCS）的NCTC - 109中培养96小时的80%大鼠囊胚（Wistar，SW72）从透明带孵化出来，并形成了滋养层巨细胞层。37%的大鼠囊胚发育出内细胞团（ICM），其中约7%由两个胚层（外胚层和内胚层）组成，而NMRI小鼠的这一比例为84%。体内已孵化的培养大鼠囊胚获得了明显更好的ICM发育。与体内情况类似，植入NCTC - 109 - FCS后的大鼠囊胚中存在乳酸脱氢酶 - 5（LDH - 5）。C57BL小鼠囊胚在NCTC - 109 - FCS中的分化与大鼠一样差。详细研究了大鼠和小鼠囊胚在NCTC - 109 - FCS中的ICM发育。由于豚鼠补体不能裂解大鼠囊胚的滋养外胚层细胞，因此使用来自不同物种（例如人、大鼠和兔补体）的补体通过免疫手术分离这两个物种的ICM。所有通过免疫手术分离的大鼠ICM在48小时内退化，但用大鼠或兔补体分离的小鼠ICM比用豚鼠补体分离的小鼠ICM发育得明显更好。对大鼠和小鼠囊胚进行囊胚总细胞数（BTCN）以及通过免疫手术分离的ICM细胞数的测定，以研究体外植入前ICM的生长动力学。在小鼠中，在NCTC - 109 - FCS中植入前的48小时以及体内植入前的16 - 24小时内，观察到BTCN和ICM细胞数均呈指数增加。在大鼠中，仅在NCTC - 109 - FCS中的前24小时发现BTCN翻倍，并且在培养的前48小时内ICM细胞数几乎没有增加。因此，NCTC - 109 - FCS中囊胚的ICM生长在小鼠植入前后受到刺激，而在大鼠中甚至在植入前就受到抑制。