Pregnancies established from water buffalo ( [formula omitted]) blastocysts derived from in vitro matured, in vitro fertilized oocytes and co-cultured with cumulus and oviductal cells

• Published in: Theriogenology Vol. 42; no. 4; pp. 591 - 600
• Main Authors: Madan, M.L., Chauhan, M.S., Singla, S.K., Manik, R.S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.1994
• Subjects: blastocysts; BUFALO DOMESTICO; buffalo; BUFFLE DOMESTIQUE; CALCIO; CALCIUM; CATION; CATIONES; CATIONS; CELLS; CELLULE; CELULAS; CULTIVO DE EMBRIONES; CULTURE D'EMBRYON; CULTURE MEDIA; CUMULUS OOPHORUS; DESARROLLO EMBRIONARIO; DEVELOPPEMENT EMBRYONNAIRE; DOMESTIC BUFFALOES; EMBRYO CULTURE; EMBRYO TRANSFER; EMBRYONIC DEVELOPMENT; EXPERIMENTACION IN VITRO; EXPERIMENTATION IN VITRO; FECONDATION; FECUNDACION; FERTILIZATION; IN VITRO EXPERIMENTATION; in vitro fertilization; in vitro maturation; INDE; INDIA; MEDIO DE CULTIVO; MILIEU DE CULTURE; Oocytes; OVA; OVIDUCTE; OVIDUCTOS; OVIDUCTS; OVULE; OVULO; TRANSFERENCIA DE EMBRIONES; TRANSFERT EMBRYONNAIRE; blastocysts; in vitro fertilization; in vitro maturation; Oocytes; buffalo
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/0093-691X(94)90376-T

Buffalo ovaries were collected immediately after slaughter and were transported to laboratory in sterile saline at 37° C. Follicular oocytes with the cumulus mass aspirated from 2 to 6 mm in diameter follicles were cultured in TCM-199 medium supplemented with 10% buffalo estrus serum (BES) in 5% CO 2 at 38.5° C. After 20 to 24 h of incubation, the oocytes were inseminated with precapacitated frozen thawed spermatozoa for 6 h. The fertilization rate was 78.15% of the matured oocytes. Over an in vitro culture period of 3 to 9 d, 4.02% of the inseminated oocytes developed to the morula stage when cultured with cumulus cells alone and 17.83% when cumulus cells plus oviductal epithelial cells were used. The percentage of developed blastocysts was very low (0.57%) when the oocytes were co-cultured with cumulus cells from the original oocytes. However, 8% of the inseminated oocytes that were denuded 3 d after insemination developed to the blastocyst stage when they were co-cultured with cumulus and oviductal epithelial cells. Sixteen early/expanded blastocysts were transferred non-surgically to 16 recipients. Four of the 16 recipients became pregnant, of which 2 delivered normal buffalo male calves.