title: In-vitro development of in-vivo produced rhesus monkey morulae and blastocysts to hatched, attached, and post-attached blastocyst stages: morphology and early secretion of chorionic gonadotrophin.
abstract: The earliest time of secretion of chorionic gonadotrophin (CG) by primate embryos and its role during preimplantation development and implantation are not clearly determined. We cultured in-vivo fertilized/developed zona-intact, morphologically normal morulae (n = 11) and early blastocysts (n = 11), freshly recovered (by non-surgical uterine flushing) on days 5 and 6 of pregnancy, respectively (day 0 = the day following LH surge), from non-superovulated naturally bred rhesus monkeys (Macaca mulatta). Embryos were cultured for a minimum of 24 days in dishes containing 1 ml of CMRL-1066 supplemented with 20% bovine fetal serum in a humidified atmosphere of 5% CO2 in air at 37 degrees C. The culture medium was changed every 48 h. The percentage of hatched blastocysts, developed from morulae and early blastocysts, was 90.9; elapsed times (mean +/- SEM) were 67.8 +/- 4.4 h (morula) and 37.8 +/- 3.6 h (blastocyst). The minimum number of Hoechst-stained cells/hatched blastocyst was 531. The mean diameter (+/- SEM) of cultured embryos increased from 180 microns at the beginning of culture to 374 +/- 28 and 450 +/- 19 microns at the fully expanded and hatched blastocyst stages, respectively. Hatched blastocysts continued to expand (maximum diameter: 1125 +/- 25 microns); after an additional 94-96 h they attached firmly to the serum-coated dishes and produced highly proliferating multinucleate trophectodermal cells, extending to a maximum diameter of 2-6 mm by 11-21 days of culture. Biologically active CG in embryo-grown, serial spent media samples was measured in a mouse Leydig cell bioassay.(ABSTRACT TRUNCATED AT 250 WORDS)
authors: Wisconsin Regional Primate Research Center, University of Wisconsin, Madison 53715-1299.
authors: P B
authors: PB
authors: Seshagiri
authors: J P
authors: JP
authors: Hearn
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000378:metabolism*; D010064:Embryo Implantation / Q000502:physiology*; D005314:Embryonic and Fetal Development / Q000502:physiology; D005260:Female; D008253:Macaca mulatta; D009028:Morula / Q000502:physiology*; D013997:Time Factors
chemical_list: D006063:Chorionic Gonadotropin