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Effects of the addition of hormone in maturation medium for in vitro production ofembryo (IVP). Hidayati, Nurhasanah; Sugiarti, Tatit; Situmorang, Polmer; Triwulanningsih, Endang; Lubis, Adriana
Indonesian Journal of Animal and Veterinary Sciences Vol 3, No 1 (1998)
Publisher : Indonesian Animal Sciences Society

A study on the effects of hormone FSH, hCG and estrogen in maturation medium on embryo production (IVP) was conducted. Ovaries of dairy cows were obtained from slaughtered house and oocytes were collected by aspiration and slicing. Oocytes were matured in TCM-199 media containing one of each hormonal treatments : 10 ug/ml FSH, 2 IU hCG, 1 ug/ml estrogen, 10 ug/ml FSH + 2 IU hCG, 10 ug/ml FSH + 1 ug/ml estrogen, and 10 ug/ml FSH + 2 IU hCG + 1 ug/ml estrogen for 24 hours. Fertilization was conducted in thyroid albumin lactate pyruvate (TALP) media containing 10 ug/ml heparin for 18-24 hours and co-cultured in synthetic oviduct fluid (Sof-media) using C02 incubator at 38oC. Every 48 hours the embryos were moved into fresh Sofmedia and embryo evaluation was done on day 7using a microscope . From a total of 293 oocytes studied resulted 60.4% of zygotes which develop to 9.4% young embryos ( cell numbers <16), 45.3% morulae and 5,7% blastocysts. Hormones used singly did not significantly affect the production of embryo . The highest mean percentages of fertilization wasobtained in FSH (73.3%) and the lowest in estrogen (59.6%), but the mean percentage of blastocyst was higher in estrogen . The mean percentages of young embryos, morulae and blastocysts were 4.8, 66.1, 2.4 ; 18 .8, 41 .6, 6.6 and 3 .3, 46.6, 10 .0 for FSH, hCG and estrogen, respectively . Using a combination of FSH with hCG and estrogen did not significantly increase the production of embryo . The mean percentages of fertilization, young embryo, morulae and blastocysts were 55.2, 9.3, 43.4 and 2.6 ; 51 .7, 4.8, 35.8 and 11 .1 and 56 .0, 12 .3, 42.3 and 1 .4 for FSH+hCG, FSH+estrogen and FSH+hCG+estrogen respectively . Â Keywords: Embryo, fertilization, hormone, in vitro

SELEKSI DAN KAPASITASI SPERMATOZOA DENGAN METODE PERCOLL GRADIENT VKYUK FERTILISASI OOSIT DAN PRODUKSIEMBRIO IN VITRO PADA SAPI Triwulanningsih, Endang; Toelihere, MR; Yusuf, TL; Purwantara, B; Diwyanto, K; Rutledge, JJ
BERITA BIOLOGI Vol 6, No 3 (2002)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

This research has been conducted at the laboratory of in vitro fertilization of the University ofWisconsin, USA. These embryos may be used for improving genetic value of Indonesian cattle. Ovaries were collected from slaughterhouse in Wisconsin. Oocytes were matured in TCM-199 medium enriched with FSH 10 ilm, estradiol 17 P lul/ml and 10 % FCS for 20 hours. The oocytes were fertilized in vitro with motile sperm selected and capacitated by using the percoll gradient with 2 ml vs 0.5 ml per layer as treatment A and B respectively. Sperm and oocytes were incubated in fertilization medium (mTALP) for 20 hours. All zygotes were cultured in CRlaa medium up to btastocyst stage and were fed with serum 5 iV 50 )j.l in culture medium on day 6. Percentages of cleavage, morula, blastocyst, expanded blastocyst, unfertilized and degenerated ova in this study were 86.3 vs 91.6 %, 53.3 % vs 75.9 %; 32.6 % vs 63.4 %; 21.1 % vs 33.0 %; 13.7 % vs 8.4 %, 32.9% vs 15.6 % for treatment A (n=1007) vs B (n=1055), respectively. Based on result of this study, it is concluded that the best method for IVP (in vitro production) of cattle embryos is using percoll gradient with 500 ul per layer.

Using CR1aa versus KSOM as the culture medium for in vitro embryo production of cattle Endang Triwulanningsih; M.R Toelihere; J.J Rutledge; T.L Yusuf; B Purwantara; K Diwyanto
Jurnal Ilmu Ternak dan Veteriner Vol 7, No 1 (2002): MARCH 2002
Publisher : Indonesian Center for Animal Research and Development (ICARD)

This research has been conducted at the laboratory of in vitro fertilization in the Department of Animal Science University of Wisconsin, USA. These embryos can be used for improving genetic value of Indonesian cattle. Oocytes were matured in TCM- 199 medium (in 5% CO2 incubator and at 390C) enriched with follicle stimulating hormone (FSH) 10 μl/ml, oestradiol 17 β 1μl/ml and 10% Fetal Calf Serum (FCS). The oocytes were fertilized in vitro with motile sperm and incubation between sperm and oocytes in fertilization medium Tyroide Albumin Lactate Pyruvate (TALP) for 20 hours. All zygotes were cultured in CR1aa (n=1549) medium versus modification of protein-free pottasium simplex optimized medium (KSOM) (n=675) up to blastocyst stage and were fed FCS 5 μl/50 μl medium on day 6, as treatment A and B respectively. Data were analyzed by completely randomized design with SAS program. Percentages of cleavage, morula, blastocyst, expanded blastocyst, unfertilized and degenerated ova in this study were 91.4% vs 75.6 %; 75.6% vs 58.9%; 61.5% vs 38.5%; 31.2% vs 5.1%, 8.6% vs 24.4%, 15.7% vs 8% which were significantly different (P<0.01) for treatment CR1aa and KSOM respectively. Based on this study, CR1aa medium is better culture medium than KSOM for efficient in vitro production (IVP) of bovine embryos. Key words: Oocytes, in vitro fertilization, embryo, blastocyst, culture medium

Effects of the addition of hormone in maturation medium for in vitro production ofembryo (IVP). Polmer Situmorang; Endang Triwulanningsih; Adriana Lubis; Nurhasanah Hidayati; Tatit Sugiarti
Jurnal Ilmu Ternak dan Veteriner Vol 3, No 1 (1998)
Publisher : Indonesian Center for Animal Research and Development (ICARD)

A study on the effects of hormone FSH, hCG and estrogen in maturation medium on embryo production (IVP) was conducted. Ovaries of dairy cows were obtained from slaughtered house and oocytes were collected by aspiration and slicing. Oocytes were matured in TCM-199 media containing one of each hormonal treatments : 10 ug/ml FSH, 2 IU hCG, 1 ug/ml estrogen, 10 ug/ml FSH + 2 IU hCG, 10 ug/ml FSH + 1 ug/ml estrogen, and 10 ug/ml FSH + 2 IU hCG + 1 ug/ml estrogen for 24 hours. Fertilization was conducted in thyroid albumin lactate pyruvate (TALP) media containing 10 ug/ml heparin for 18-24 hours and co-cultured in synthetic oviduct fluid (Sof-media) using C02 incubator at 38oC. Every 48 hours the embryos were moved into fresh Sofmedia and embryo evaluation was done on day 7using a microscope . From a total of 293 oocytes studied resulted 60.4% of zygotes which develop to 9.4% young embryos ( cell numbers <16), 45.3% morulae and 5,7% blastocysts. Hormones used singly did not significantly affect the production of embryo . The highest mean percentages of fertilization wasobtained in FSH (73.3%) and the lowest in estrogen (59.6%), but the mean percentage of blastocyst was higher in estrogen . The mean percentages of young embryos, morulae and blastocysts were 4.8, 66.1, 2.4 ; 18 .8, 41 .6, 6.6 and 3 .3, 46.6, 10 .0 for FSH, hCG and estrogen, respectively . Using a combination of FSH with hCG and estrogen did not significantly increase the production of embryo . The mean percentages of fertilization, young embryo, morulae and blastocysts were 55.2, 9.3, 43.4 and 2.6 ; 51 .7, 4.8, 35.8 and 11 .1 and 56 .0, 12 .3, 42.3 and 1 .4 for FSH+hCG, FSH+estrogen and FSH+hCG+estrogen respectively . Keywords: Embryo, fertilization, hormone, in vitro

SELEKSI DAN KAPASITASI SPERMATOZOA DENGAN METODE PERCOLL GRADIENT VKYUK FERTILISASI OOSIT DAN PRODUKSIEMBRIO IN VITRO PADA SAPI Endang Triwulanningsih; MR Toelihere; TL Yusuf; B Purwantara; K Diwyanto; JJ Rutledge
BERITA BIOLOGI Vol 6, No 3 (2002)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v6i3.1214

This research has been conducted at the laboratory of in vitro fertilization of the University ofWisconsin, USA. These embryos may be used for improving genetic value of Indonesian cattle. Ovaries were collected from slaughterhouse in Wisconsin. Oocytes were matured in TCM-199 medium enriched with FSH 10 \i\lm\, estradiol 17 P lul/ml and 10 % FCS for 20 hours. The oocytes were fertilized in vitro with motile sperm selected and capacitated by using the percoll gradient with 2 ml vs 0.5 ml per layer as treatment A and B respectively. Sperm and oocytes were incubated in fertilization medium (mTALP) for 20 hours. All zygotes were cultured in CRlaa medium up to btastocyst stage and were fed with serum 5 \iV 50 )j.l in culture medium on day 6. Percentages of cleavage, morula, blastocyst, expanded blastocyst, unfertilized and degenerated ova in this study were 86.3 vs 91.6 %, 53.3 % vs 75.9 %; 32.6 % vs 63.4 %; 21.1 % vs 33.0 %; 13.7 % vs 8.4 %, 32.9% vs 15.6 % for treatment A (n=1007) vs B (n=1055), respectively. Based on result of this study, it is concluded that the best method for IVP (in vitro production) of cattle embryos is using percoll gradient with 500 ul per layer.

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