Ultrastructural Morphometry of Bovine Compact Morulae Produced In Vivo or In Vitro

• Published in: Biology of reproduction 2000-05, Vol.62 (5), p.1459-1465
• Main Authors: Crosier, A E, Farin, P W, Dykstra, M J, Alexander, J E, Farin, C E
• Format: Article
• Language: English
• Subjects: Animals; Cattle; Cell Culture Techniques - methods; Cytoplasm; Female; Fertilization in Vitro; Morula - cytology; Morula - ultrastructure
• ISSN: 0006-3363; 1529-7268
• DOI: 10.1095/biolreprod62.5.1459

The objective of this study was to compare the ultrastructure of bovine compact morulae produced in vivo or in vitro using morphometric analysis. Compact morulae produced in vivo were obtained from superovulated Holstein cows. Compact morulae produced in vitro were obtained from cumulus-oocyte complexes aspirated from ovaries of Holstein cows. The complexes were matured and fertilized in vitro. At 20 h postinsemination (hpi), zygotes were distributed into 1 of 3 culture media: 1) IVPS (in vitro produced with serum): TCM-199 + 10% estrous cow serum (ECS); 2) IVPSR (in vitro produced with serum restriction): TCM-199 + 1% BSA until 72 hpi followed by TCM-199 + 10% ECS from 72 to 144 hpi; 3) mSOF (modified synthetic oviductal fluid): SOF + 0.6% BSA. At 144 hpi, five grade 1 compact morulae from each of the four treatments were prepared for transmission electron microscopy. The volume density occupied by cellular components was determined by the point-count method using a sampling of seven to nine random micrographs from each compact morula. The volume density of lipid was greater ( P < 0.05) in compact morulae from IVPS, IVPSR, and mSOF treatments compared with those produced in vivo. There was a reduced proportional volume of total mitochondria in compact morulae from the IVPS treatment compared with those produced in vivo ( P < 0.05). For compact morulae from the IVPS culture treatment, the volume density of vacuoles was greater than that for compact morulae produced in vivo ( P < 0.05). The cytoplasmic-to-nuclear ratio for compact morulae from the IVPS treatment was increased ( P < 0.05) compared with the ratio for those produced in vivo. In conclusion, compact morulae produced in vitro differed ultrastructurally from those produced in vivo. Compact morulae produced in IVPS culture medium possessed the greatest deviations in cellular ultrastructure.