Development and quality of bovine embryos produced in vitro using growth factor supplemented serum-free system  

作　　者：Arindam Dhali Vahida M. Anchamparuthy Steve P. Butler Isis K. Mullarky Ronald E. Pearson Frank C. Gwazdauskas 

机构地区：[1]不详

出　　处：《Open Journal of Animal Sciences》2011年第3期97-105,共9页动物科学期刊（英文）

摘　　要：The influence of a growth factor supplemented serum-free system on the development, gene expression, and cryotolerance of in vitro pro- duced bovine embryos was investigated. To assess the embryo development and gene ex- pression in blastocysts, abattoir-derived oo- cytes (obtained from 3 - 10 or <3 mm follicles) were matured and fertilized in serum-free media and cultured in synthetic oviductal fluid sup- plemented with fetal bovine serum (FBS, 4%), epidermal growth factor (EGF, 10 ng/mL), insulin like growth factor-1 (IGF-1, 100 ng/mL), stem cell factor (SCF, 50 ng/mL) or combinations of the growth factors. Expressions of selected gene transcripts were relatively quantified in the d 8 blastocysts. To assess the cryotolerance, d 4 morulae (derived from 3 - 10 mm follicles and cultured with the supplementation of FBS or combinations of the growth factors) were vitri- fied, thawed and cultured (with respective sup- plementations). Total cell number and DNA frag- mentation in blastocysts derived from the vitri- fied morulae were assessed through TUNEL assay. The rate (%) of cleavage, blastocyst and expanded/hatched blastocyst did not differ among the culture medium supplementations within the follicle size of 3 - 10 mm (range 65.1 ± 4.3 - 75.4 ± 3.9;22.4 ± 3.9 - 36.4 ± 3.6;and 11.2 ± 2.9 - 23.3 ± 3.2, respectively) or <3 mm (range 59.3 ± 4.2 - 74.5 ± 3.7;15.0 ± 3.5 - 28.7 ± 4.5;and 9.3 ± 2.8 - 17.3 ± 2.7, respectively). Nevertheless, significantly lower (P < 0.05) cleavage and blastocyst rates with FBS and lower blastocyst rate with SCF supplementations were observed for the oocytes derived from <3 compared to 3 - 10 mm follicles. The expression patterns of BCL-2, BAX, HSP1A1, GJA1 and BIRC5 tran- scripts varied significantly (P < 0.05) in all cases, except for BIRC5 in the blastocysts derived from 3 - 10 mm follicles. Following thaw and culture, the development (%) of vitrified morulae into expanded/hatched blastocysts was lower (P < 0.01) with the supplementation of growth fac- tors compared to FBS. IThe influence of a growth factor supplemented serum-free system on the development, gene expression, and cryotolerance of in vitro pro- duced bovine embryos was investigated. To assess the embryo development and gene ex- pression in blastocysts, abattoir-derived oo- cytes (obtained from 3 - 10 or <3 mm follicles) were matured and fertilized in serum-free media and cultured in synthetic oviductal fluid sup- plemented with fetal bovine serum (FBS, 4%), epidermal growth factor (EGF, 10 ng/mL), insulin like growth factor-1 (IGF-1, 100 ng/mL), stem cell factor (SCF, 50 ng/mL) or combinations of the growth factors. Expressions of selected gene transcripts were relatively quantified in the d 8 blastocysts. To assess the cryotolerance, d 4 morulae (derived from 3 - 10 mm follicles and cultured with the supplementation of FBS or combinations of the growth factors) were vitri- fied, thawed and cultured (with respective sup- plementations). Total cell number and DNA frag- mentation in blastocysts derived from the vitri- fied morulae were assessed through TUNEL assay. The rate (%) of cleavage, blastocyst and expanded/hatched blastocyst did not differ among the culture medium supplementations within the follicle size of 3 - 10 mm (range 65.1 ± 4.3 - 75.4 ± 3.9;22.4 ± 3.9 - 36.4 ± 3.6;and 11.2 ± 2.9 - 23.3 ± 3.2, respectively) or <3 mm (range 59.3 ± 4.2 - 74.5 ± 3.7;15.0 ± 3.5 - 28.7 ± 4.5;and 9.3 ± 2.8 - 17.3 ± 2.7, respectively). Nevertheless, significantly lower (P < 0.05) cleavage and blastocyst rates with FBS and lower blastocyst rate with SCF supplementations were observed for the oocytes derived from <3 compared to 3 - 10 mm follicles. The expression patterns of BCL-2, BAX, HSP1A1, GJA1 and BIRC5 tran- scripts varied significantly (P < 0.05) in all cases, except for BIRC5 in the blastocysts derived from 3 - 10 mm follicles. Following thaw and culture, the development (%) of vitrified morulae into expanded/hatched blastocysts was lower (P < 0.01) with the supplementation of growth fac- tors compared to FBS. I

关 键 词：IVF Growth Factors EMBRYO DEVELOPMENT Gene Expression CRYOTOLERANCE BOVINE 

分 类 号：R73[医药卫生—肿瘤]