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In vitro co-culture of in vitro fertilized (IVF) bovine embryos with different cell monolayers
THERIOGENOLOGY IN VITRO CO-CULTURE OF IN VITRO FBRTILISED (IVF) BOVINE EMBRYOS WITH DIFFBRBNT CELL MONOLAYERS ahao
Eiaoxia,
Jin
Biotech. Chinese
zhangun, Jiang Shie, Zhu Yuding
Liu
Jianmin,
of Animal Science, Lab., Institute Academy of Agricultural Sciences, Beijing 100094, P.R. China
The objective of this study was to compare the efficiency The whole experiment was CO-Culture systems. performed under 5% CO*, at 39OC. Early embryos of BIVF cocultured either with bovine oviduct epithelium monolayer (BOEM) or with bovine granular cell monolayer (BGCM) had both improved viability and showed higher cleavage rate than embryos cultured in medium alone (TCM 199 + 10% FCS (SIGMA, (67.8% (61/90), 63.0% (85/135) and 27.10% USA)), CC, (29/107), respectively). co-culture of embryos with BGCM resulted in more 4-8 cell 42185) when embryos cultured 48 h after (49.5%, fertilisation than those with BOBM (18.0% 11/61); but, more cleaved embryos co-cultured with BOEM passed the 8-16 cell block (90.2%, 45/61) than those with BGCM (60.0%, 52185) when cultured for 72-96 h. overall, co-culture of embryos with BOEM can produce more non-surgically transferable embryos (morulae and blastocysts) (30.3%, 27/89) than those with BGCM (20.18%, 36/170). Bmbryos co-cultured with BOEM still lagged behind in development compared with embryos flushed from superovulated females. To improve the effect of the co-culture systems, early embryos of BIVF were at first co-cultured with BGCM for about 48 h, then cleaved embryos were moved to the BOEM coculture system for further culture. At the end of 48h, embryos still had the high 4-8 cell rate (59%, 26- /44), at 72-96 h more cleaved embryos passed the 8-16 cell block more high (77%, 20/26), and after 168-192 h of co-culture, quality morulae and blastocysts (38%, 17144) were obtained. Moreover, the speed of embryo development (cleaving, expanding and hatching) in vitro was similar to that in vivo. We conclude the (1) granular cells can support bovine in vitro fertiliaed eggs to cleave normally to the 4-8 cell stage, (2) oviduct cells play a specific role in supporting passage of embryos through the 8-16 cell block and (3) coculture of BIVF embryos with different cells at different embryo stages can be a convincing way to produce nonsurgically transferable embryos , and to manipulate embryos, such as gene transfer and embryo sexing, etc. while preserving the embryo viability.
of different
JANUARY
1991 VOL. 35 NO. 1
295
Eiaoxia,
Jin
Biotech. Chinese
zhangun, Jiang Shie, Zhu Yuding
Liu
Jianmin,
of Animal Science, Lab., Institute Academy of Agricultural Sciences, Beijing 100094, P.R. China
The objective of this study was to compare the efficiency The whole experiment was CO-Culture systems. performed under 5% CO*, at 39OC. Early embryos of BIVF cocultured either with bovine oviduct epithelium monolayer (BOEM) or with bovine granular cell monolayer (BGCM) had both improved viability and showed higher cleavage rate than embryos cultured in medium alone (TCM 199 + 10% FCS (SIGMA, (67.8% (61/90), 63.0% (85/135) and 27.10% USA)), CC, (29/107), respectively). co-culture of embryos with BGCM resulted in more 4-8 cell 42185) when embryos cultured 48 h after (49.5%, fertilisation than those with BOBM (18.0% 11/61); but, more cleaved embryos co-cultured with BOEM passed the 8-16 cell block (90.2%, 45/61) than those with BGCM (60.0%, 52185) when cultured for 72-96 h. overall, co-culture of embryos with BOEM can produce more non-surgically transferable embryos (morulae and blastocysts) (30.3%, 27/89) than those with BGCM (20.18%, 36/170). Bmbryos co-cultured with BOEM still lagged behind in development compared with embryos flushed from superovulated females. To improve the effect of the co-culture systems, early embryos of BIVF were at first co-cultured with BGCM for about 48 h, then cleaved embryos were moved to the BOEM coculture system for further culture. At the end of 48h, embryos still had the high 4-8 cell rate (59%, 26- /44), at 72-96 h more cleaved embryos passed the 8-16 cell block more high (77%, 20/26), and after 168-192 h of co-culture, quality morulae and blastocysts (38%, 17144) were obtained. Moreover, the speed of embryo development (cleaving, expanding and hatching) in vitro was similar to that in vivo. We conclude the (1) granular cells can support bovine in vitro fertiliaed eggs to cleave normally to the 4-8 cell stage, (2) oviduct cells play a specific role in supporting passage of embryos through the 8-16 cell block and (3) coculture of BIVF embryos with different cells at different embryo stages can be a convincing way to produce nonsurgically transferable embryos , and to manipulate embryos, such as gene transfer and embryo sexing, etc. while preserving the embryo viability.
of different
JANUARY
1991 VOL. 35 NO. 1
295