+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

In vitro maturation of bovine cumulus enclosed primary oocytes and their subsequent in vitro fertilization and cleavage



In vitro maturation of bovine cumulus enclosed primary oocytes and their subsequent in vitro fertilization and cleavage



Journal of Dairy Science 68(6): 1456-1462



Cumulus enclosed primary oocytes from 2 to 4-mm bovine follicles were matured in vitro in Minimum Essential Medium containing follicle-stimulating hormone (0, .1, 1, 10, 50, or 100 micrograms/ml) or human chorionic gonadotropin (0, .1, 1, or 10 IU/ml) for 48 h at 37 degrees C under paraffin oil. Cumulus mass expansion comparable to that seen in vivo occurred in 18% of the control oocytes, 39% of those cultured in human chorionic gonadotropin, and 56% of those cultured in follicle-stimulating hormone. The optimum follicle-stimulating hormone concentration for cumulus expansion was 1 microgram/ml, and this was then used to mature oocytes individually or in groups of 5 for in vitro fertilization. Ejaculated bovine semen, extended 1:10 with yolk-TES-Tris extender and stored 24 to 48 h at 4 degrees C, was warmed, washed once with Minimum Essential Medium, and 500,000 motile sperm/ml were used to inseminate the matured oocyte-cumulus cell complexes. Criteria for fertilization was cleavage to the two-cell stage 48 h after insemination. Oocytes, inseminated individually, cleaved with a frequency of 5%, whereas 15% of those inseminated in groups of 5 cleaved, perhaps as the result of cumulus factors enhancing capacitation. The cleavage rate for the parthenogenetic control with killed spermatozoa was 0%. Therefore, primary oocytes matured in vitro to secondary oocytes were successfully fertilized in vitro and cleaved to at least the two-cell stage in the Minimum Essential Medium. Individual differences between bulls in ability to fertilize in vitro were noted.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 001387179

Download citation: RISBibTeXText

PMID: 3926843

DOI: 10.3168/jds.s0022-0302(85)80983-8


Related references

Effect of cumulus cell removal of in vitro matured bovine oocytes prior to in vitro fertilization on subsequent cleavage rate. Theriogenology 57(4): 1347-1355, 2002

Apoptosis in cumulus cells during in vitro maturation of bovine cumulus-enclosed oocytes. Reproduction 125(3): 369-376, 2003

Studies on in vitro maturation and fertilization of bovine oocytes. II. Effects of some reproductive hormones on the maturation, fertilization and the subsequent development potential of bovine oocytes in vitro. Memoirs of the College of Agriculture, National Taiwan University 29(1): 150-160, 1989

Increasing the cAMP concentration during in vitro maturation of pig oocytes improves cumulus maturation and subsequent fertilization in vitro. Theriogenology 83(3): 344-352, 2015

Paracrine factors from cumulus-enclosed oocytes ensure the successful maturation and fertilization in vitro of denuded oocytes in the cat model. Fertility and Sterility 91(5 Suppl): 2051-2060, 2009

Studies on in vitro maturation of bovine follicular oocytes. 2. Effects of oestrous bovine serum, bovine follicular fluid and matured cumulus cells on in vitro maturation and fertilization of follicular oocytes. Korean Journal of Dairy Science 11(3): 139-146, 1989

Influence of co-culture with denuded oocytes during in vitro maturation on fertilization and developmental competence of cumulus-enclosed porcine oocytes in a defined system. Animal Science Journal 87(4): 503-510, 2016

In vitro fertilization and cleavage capability of bovine follicular oocytes classified by cumulus cells and matured in vitro. Theriogenology 30(3): 489-496, 1988

Influence of cumulus cells on in vitro fertilization of bovine oocytes derived from in vitro maturation. Animal Reproduction Science 38(1-2): 37-48, 1995

In vitro production of bovine blastocysts by in vitro maturation and fertilization of oocytes and subsequent in vitro culture. Zuchthygiene 24(3): 134-139, 1989

Cumulus expansion in vitro fertilization and embryonic development after in vitro maturation of bovine oocytes in the presence of follicle stimulating or luteinizing hormone. Reproduction in Domestic Animals 25(1): 3-13, 1990

Increased glutamine metabolism in bovine cumulus cell-enclosed and denuded oocytes after in vitro maturation with luteinizing hormone. Biology of Reproduction 48(4): 815-820, 1993

Manganese inhibits in vitro nuclear maturation in cumulus-enclosed bovine oocytes through the cAMP/protein kinase A pathway. Journal of Dairy Science 85(Suppl. 1): 302-303, 2002

Role of cumulus cells and serum on the in vitro maturation, fertilization, and subsequent development of rat oocytes. Biology of Reproduction 40(4): 720-728, 1989

Timing of in-vitro fertilization of cumulus-free and cumulus-enclosed human oocytes. Human Reproduction 1(4): 237-242, 1986