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Effects of rabbit antiserum on mouse embryonic stem cells at 8-cell embryo development stage
HU Chun-chao CUI Li-fang WANG Peng-bo WANG Mian-juan DI Ke-qian LI Xiang-yun*
Acta Anatomica Sinica ›› 2013, Vol. 44 ›› Issue (6) : 812-818.
Effects of rabbit antiserum on mouse embryonic stem cells at 8-cell embryo development stage
Objective Mouse 8-cell embryos were cultured in KSOM media with rabbit antiserum to mouse embryonic stem (ES) cells, and the effect of the antiserum on mouse embryos was observed. Methods The antiserum was diluted to 1/50, 1/100 and 1/200 by KSOM medium, respectively. Results The result of morphology changes showed that the embryonic development was inhibited or even stopped in the 1/50 dilution group, while the compaction was blocked, the embryos showed fluid vacuoles in the blastula stage in the 1/100 dilution group; there was no obvious effect in the 1/200 dilution group. The treatment group with normal rabbit serum did not appear differences from the control group. Our data indicated that the compaction was blocked by antiserum; when the compacted embryos were exposed in the antiserum, decompaction occurred; prolonged decompaction beyond the 32-cell embryo resulted in an increasing proportion of malformed blastocysts. The embryo development was further tested in the 1/100 dilution group by compaction rate, blastocyst rate, inner cell mass (ICM) outgrowth rate, cell counting, embryo transplantation and fluorescent staining. The result showed that the 1/100 dilution antiserum inhibited ICM formation, decreased embryonic cell number, and creased abnormal blastocyst rate. The treated blastocysts were transferred into uterine horns and did not develop to term. Conclusion Our data suggests that the rabbit antiserum to mouse ES cells inhibits embryo compaction, ICM formation and postimplantation development.
Rabbit antiserum / Embryonic stem cell / Inner cell mass / Compaction / Culture in vitro / Mouse
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