In vitro development of rabbit pronuclear embryos in rabbit peritoneal fluid
1991
Collas, P. | Duby, R.T. | Robl, J.M.
The use of rabbit peritoneal fluid (PF) for the culture of rabbit embryos in vitro was evaluated. Development of zygotes cultured in PF and Earle's balanced salts solution (EBSS) + 10% fetal calf serum (EBSS/FCS) was compared. The effects of increasing the concentration of PF in EBSS and of culturing embryos in fractionated PF were also investigated. In addition, embryonic development in PF was compared to that in vivo. Development to hatching blastocysts was enhanced with PF (73%) compared to EBSS/FCS (3%, p < 0.001). PF manifested greater mitogenic activity than EBSS/FCS, as indicated by higher cell number in embryos at 48, 72, and 96 h post-mating/hCG (p < 0.001). PF also promoted blastocyst cell proliferation in a dose-dependent manner (r = 0.98, p < 0.01); however, embryo growth remained slower than in vivo. Culture in the high (> 30 000 Da) molecular mass fraction of PF reduced incidence of hatching (56% vs. 92%, p < 0.001) and mean cell number in Day 4 blastocysts (151 +/- 4 vs. 243 +/- 5, p < 0.001). Rates of blastocyst hatching (10%) and cell number (110 +/- 3) were further reduced in the low (< 30 000 Da) molecular mass fraction. When the high molecular mass fraction was dialyzed, embryos did not develop beyond the early morula stage. This suggests that the interaction or the synergy of high and low molecular mass components of PF is necessary for optimum development of rabbit embryos.
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