Effects of bovine serum albumin and estrous cow serum on development and ultrastructure of in vitro-produced porcine embryos.
This study evaluated the effects of bovine serum albumin (BSA; 4 mg/ml) and estrous cow serum (ECS; 10%) in North Carolina State University (NCSU) 23 medium on the development of in vitro-matured and in vitro-fertilized porcine oocytes. Early cleavage rate was significantly (P < 0.05) higher in NCSU/ECS (71.3 +/- 14.7%) vs. NCSU/BSA (60.6 +/- 4.7%). Cleavage beyond the four-cell stage was not different between the two culture media (43.5 +/- 9.5% and 41.4 +/- 17.7%, respectively). The proportion of development to blastocysts was--with borderline significance (P = 0.05)--higher in NCSU/BSA (28.0 +/- 4.4%) than in NCSU/ECS (20.4 +/- 7.3%). Blastocysts produced in NCSU/BSA had significantly (P < 0.001) higher cell numbers than those cultured in NCSU/ECS (29.5 +/- 20.1 vs. 16.9 +/- 10.8). The ultrastructure of in vitro-produced blastocysts from both culture systems was compared vs. in vivo-derived blastocysts. The latter showed a clear differentiation between trophectoderm (TE) and inner cell mass (ICM) cells. The TE cells were anchored to other TE cells or ICM cells by long, well-developed junctional complexes. The apical membrane of trophoblast cells was covered with numerous microvilli. Mitochondria were abundant, round to elongated in shape, and showed clear transverse cristae. The ultrastructure of blastocysts cultured in NCSU/BSA mimicked that of in vivo-derived embryos closely. In contrast, blastocysts from the NCSU/ECS culture system displayed an irregular ultrastructure with reduced numbers of organelles and numerous cytoplasmic inclusions, such as lipid-yolk-vacuoles and vacuoles with lipid content. In some sections of these embryos, cellular debris was detected in cytoplasm. The shape of mitochondria was more ovoid and cristae were not visible. In summary, our results demonstrate a beneficial influence of ECS in the culture medium on initial cleavage of in vitro-produced porcine embryos. Clearly negative effects of ECS in the subsequent culture period are associated with marked ultrastructural changes of embryonic cells.
Ott
Monika
M
Department of Molecular Animal Breeding and Biotechnology, Hackerstrasse 27, Oberschleissheim, Germany.
Schernthaner
Wolfgang
W
Sinowatz
Fred
F
Wolf
Eckhard
E
D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000648:ultrastructure; D002417:Cattle; D003470:Culture Media; D005314:Embryonic and Fetal Development / Q000187:drug effects / Q000502:physiology*; D004971:Estrus / Q000097:blood*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D009028:Morula / Q000187:drug effects / Q000648:ultrastructure; D009865:Oocytes / Q000187:drug effects / Q000648:ultrastructure*; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D013552:Swine / Q000196:embryology*
D003470:Culture Media; D012710:Serum Albumin, Bovine