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Journal : HAYATI Journal of Biosciences

Number and Quality of Oocytes Collected from Heterotopic Autografted Mice Ovary after PMSG Induction NURBARIAH, .; DJUWITA, ITA; MOHAMAD, KUSDIANTORO; SUPRIATNA, IMAN
HAYATI Journal of Biosciences Vol 18, No 4 (2011): December 2011
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Heterotopic grafting sites can be useful in producing oocytes for in vitro Fertilization, therefore, maximising the oocyte yield from the graft by gonadotrophin stimulation would be advantageous. The aim of this study was to investigate the number and quality of oocytes collected from heterotopic autografted ovary after Pregnant Mare Serum Gonadothropin (PMSG) induction. Graft recipients were treated either with or without PMSG stimulation 48 hours prior to graft collection. Ovarian tissue from four weeks old mice (DDY strain) were autotransplanted under the kidney capsule of the same ovariectomized mice and the oocytes were collected 21 days after autotransplantation. The results showed that the average number of oocytes collected from autografted ovaries without PMSG induction were 9.0. + 2.8 not significantly different with those received PMSG induction, 10.9 + 5.1. The percentage of matured and fertilized oocytes and the developed embryos from the autografted ovaries without PMSG induction were 52.4, 33.4, and 26.0%, respectively not significantly different with those received PMSG induction, 53.2, 35.1, and 29.9%, respectively. The number of oocytes and the capacity to matured, fertilized and developed were significantly lower (P < 0.05) compared to the superovulated nongrafted (control) ovaries. In conclusion, PMSG induction on the graft recipients did not significantly increase oocytes yield from grafted heterotopic ovaries. The number and quality of oocytes produced from the autografted ovaries were lower than the superovulated nongrafted ovaries, but still can be used for in vitro embryo production after sequential in vitro maturation and fertilization.
Development of Domestic Cat Embryo Produced by Preserved Sperms ERIANI, KARTINI; BOEDIONO, ARIEF; DJUWITA, ITA; SUMARSONO, SONY HERU; AL-AZHAR, AL-AZHAR
HAYATI Journal of Biosciences Vol 15, No 4 (2008): December 2008
Publisher : Bogor Agricultural University, Indonesia

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Abstract

The ability to mature and fertilize oocytes of endangered species may allow us to sustain genetic and global biodiversity. Epididymis sperms may be the last chance to ensure preservation of genetic materials after injury or death of a valuable animal. Studies have been conducted to determine wether both epididymis sperms and oocytes can be used to produce viable embryos and offspring. The purpose of this study was to determine how long cats sperms contained in epididymis were remain motile and had intact membranes when preserved at 4 oC, and to determine whether such those preserved sperms are able to fertilize oocytes. Epididymis was preserved immediately in phosphate buffer saline at 4 oC for 1, 3, and 6 days. The observation of sperm quality and viability after preservation was performed by vital staining acrosom and Hoechst-Propidium Iodine. Biological functions of sperms were evaluated by in vitro culture technique for fertilization, micro fertilization and embryonic development rate in CR1aa medium. The results showed that average motility of sperms collected from ductus deferens, cauda and corpus epididymis decreased not significantly (P > 0.05) from 0, 1, 3, and 6 days of preservation times (from 83.0%, 80.2%, 79.0%; 80.9%, 75.0%, 75.5%; 52.0%, 63.2%, 55.0% to 34.6%, 34.6%, 33.3%, respectively). The general results showed that sperms from epididymis preserved for 1, 3, and 6 days can be used for IVF. The rate of embryonal cleavage produced by IVF technique using sperms collected from epididymis preserved for 1-, 3- and 6-days were 33.3, 26.7, and 20.0%, respectively and significantly different (P < 0.05) from that of controll (50.0%). In conclusion, sperms contained in epididyimis preserved at 4 oC in PBS (Phospate Buffer Saline) for 1-6 days can be used to IVF and in vitro production of cat embryos. Key words: gamet, preservation, in vitro fertilization
In Vitro Fertilization and Embryo Development DJUWITA, ITA; BOEDIONO, ARIEF; AGUNGPRIYONO, SRIHADI; SUPRIATNA, IMAN
HAYATI Journal of Biosciences Vol 12, No 2 (2005): June 2005
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Experiments were conducted on the morphology, fertilization and embryo development rate of vitrified ovine oocytes matured in vitro. Three vitrification solutions were used for vitrification. PBS supplemented with 1% BSA, 30% ethylene glycol was added by one of three different sucrose concentrations, 1.00 M (VS1), 0.50 M (VS2), and 0.25 M (VS3). The results showed that the percentages of normal vitrified oocytes after warming were 78 and 63% in VS1 and VS2, respectively, which was significantly higher as compared for VS3. The fertilization rates were 59 and 66% in VS1 and VS2, respectively, which were also significantly higher as compared with VS3 (35%). Zygote viability after 18 h was 57; 43; and 40%, for VS1,VS2, and VS3, respectively, which was not significantly different. The incidence of polyspermic penetration increased with increasing sucrose concentration, i.e 23, 11, and 9% in VS1, VS2, and VS3, respectively, as compared with unvitrified oocytes (4%). The cleavage rate of vitrified oocytes in VS1 was 13.2% which was significantly lower (p