In vitro fertilization of bovine follicular oocytes and their development up to hatched blastocysts in vitro.

概要

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The objective of this study was to determine whether bovine follicular cells cocultured would enhance the in vitro development of bovine embryos fertilized in vitro. Bovine oocytes were matured in vitro for 20-24 hr. The medium used for matu-ration was TCM 199 (25 mM HEPES, Earle-salt) supplemented with 5% calf serum and antibiotics. Frozen epididymal spermatozoa were thawed in hot water (32-35°C) followed by washing three times with B.O. solution (Brackett and Oliphant, 1975) supplemented with 5 mM caffein. Then, the spermatozoa were preincubated for 2-3 hr in B.O. solution containing bovine serum albumin (5 mg/ml) and caffein (2.5 mM) before insemination. Six hours after insemination, the eggs were transferred to the medium for development. The medium used was TCM 199 supplemented with calf serum (0-10%). Fortyeight or 72-77 hr after fertilization, follicular cells surrounding eggs were removed but the follicular cells attached to the culture dish were not removed. The eggs were then cultured on the follicular cells in the dish for further development. The percentages of embryos developed to morula, blastocyst and hatched blastocyst were 18.5-30.4, 4.1-25.4 and 2. 1-3. 7%, respectively. The results of this study indicate that follicular cells enhance in vitro development of bovine embryos.
日本繁殖生物学会の論文