Articles

Isolation and Identification of Transforming Growth Factor β from In Vitro Matured Cumulus Oocyte Complexes WIDJIATI, .; BOEDIONO, ARIEF; SUMITRO, SUTIMAN BAMBANG; HINTING, AUCKY; AULANI’AM, .; SUSILOWATI, TRINIL
HAYATI Journal of Biosciences Vol 19, No 1 (2012): March 2012
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (92.482 KB) | DOI: 10.4308/hjb.19.1.6

Abstract

Transforming growth factor-β (TGF-β) is a two-chain polypeptide with molecular weight of 25 kDa which takes significant role in the steroidogenesis process. In the ovarian oocyte in particular, TGF-β has an important role in regulating reproductive function. TGF-β represents a key intrafollicular protein that regulates follicle development and aromatization process. The purpose of this research was to characterize and identify a protein fraction of TGF-β from the bovine isolated oocytes, which is synthesized during in vitro oocyte maturation process. Oocytes were collected from follicles with diameter of 3-8 mm. Oocytes were then matured in TCM 199 media supplemented with 5 μg/mg LH, 3% BSA, and 50 μg/ml gentamicin sulfate, and cultured in CO2 incubator (5%, 38.5 oC) for 20 hours. TGF-β receptors were identified immunohistochemically. Characteristics of the TGF-β protein were determined using SDS PAGE and TGF-β specification was tested using Western Blotting. The results showed that TGF-β receptors were identified and found in cumulus oocyte complexes (COCs). TGF-β protein was isolated from bovine oocytes with molecular weight 25 kDa and it was identified by Western blotting methods in the same molecular weight.
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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (90.643 KB) | DOI: 10.4308/hjb.15.4.155

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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (86.772 KB) | DOI: 10.4308/hjb.12.2.73

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
Maturation Rate of Ovine Oocytes from Different Reproductive Status and Maturation Medium BOEDIONO, ARIEF; YULNAWATI, YULNAWATI; SETIADI, MOHAMAD AGUS
HAYATI Journal of Biosciences Vol 13, No 4 (2006): December 2006
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (68.228 KB) | DOI: 10.4308/hjb.13.4.131

Abstract

The aim of the present study was to determine the number of follicles, oocyte quality and maturation rate of oocytes from pairs of ovary with different reproductive status in two maturation medium, TCM-199 as control and CR1aa as treatment. The pairs of ovary were classified into four groups: (i) ovaries with corpus luteum (CL) and dominant follicle (DF), (ii) ovaries with CL, without DF, (iii) ovaries with DF, without CL, (iv) ovaries without both CL and DF. Results of the experiment revealed that the greatest number of follicles was observed from ovary with CL without DF (15.88 + 10.68), although not significantly different (P > 0.05) with other status of ovaries. The lowest number (P < 0.05) of A grade oocytes was found from ovary with DF without CL (1.20 + 1.10). The percentage of Metaphase II was highest in TCM-199 (75.51%) with oocytes from ovaries with CL and DF, and the lowest with oocytes from ovaries with DF without CL in TCM-199 and CR1aa (42.86 and 30.95%). The study suggested that the number of oocytes with A grade were influenced by the reproductive status of ovaries. The maturation rate of A grade oocytes was influenced by the quality of oocytes and the composition of maturation medium. Key words: reproductive status, corpus luteum, dominant follicles, TCM-199, CR1aa
Koleksi Sel Telur dengan Teknik Laparoskopi untuk Produksi Embrio dan Transfer Embrio pada Domba Setiadi, Mohamad Agus; Supriatna, Iman; Boediono, Arief
Jurnal Ilmu Pertanian Indonesia Vol 12, No 2 (2007): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

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Abstract

An experiment was carried out to analyze the application of laparoscopic technique for oocyte collection, in vitro embryo production and embryo transfer in sheep. The first experiment was conducted to observe effect of gonadotropin stimulation on follicle development and laparoscopic technique for oocytes aspiration. In the second experiment, effect of culture system on the embryo development in vitro was assessed and in the third experiment, the application of laparoscopic for embryo transfer has been conducted. The result showed that single dose of gonadotrophin was sufficient to support follicle development significantly and it could help follicle visualization. It also showed that laparoscopic ovum-pick up could be conducted weekly without any restriction The second series experiment showed CR1aa culture system was better than TCM 199 (29.90°/o vs 8.00%) and the changing of media was required to ensure better metabolism process for embryos. The third experiment revealed that embryo transfer could be conducted with an aid from laparoscope. In conclusion, single dose PMSG stimulation is sufficient to support follicle development for /aparoscopic ovum-pick up, the culture media changing affects the successful rate of in vitro embryo production (8% vs 25.66%) and the laparoscopy technique can be used safely for embryo transfer on sheep.Keyword: laparoscopic, oocyte, embryo transfer, sheep
DINAMIKA FOLIKEL OVARIUM DOMBA PASCATRANSPLANTASI INTRAUTERIN PADA KELINCI PSEUDOPREGNANST Sumarmin, Ramadhan; Winarto, Adi; Yusuf, Tutty Laswardi; Boediono, Arief
Majalah Ilmiah Peternakan Vol 10, No 3 (2007)
Publisher : Majalah Ilmiah Peternakan

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Abstract

ABSTRAK Tujuan penelitian ini adalah untuk mengevaluasi dinamika folikel pada ovarium domba pascatransplantasi secara intrauterin pada kelinci pseudopregnansi. Transplantasi dilakukan pada kelinci pseudopregnant hari ke 1 atau ke 7. Ovarium kembali diambil pada hari ke 5, 7, atau 9 setelah transplantasi. Untuk menentukan dinamika folikel pada ovarium domba pascatransplantasi dan menghitung jumlah folikel pada berbagai tahap perkembangan, ovarium domba pascatransplantasi dijadikan preparat histologis dengan metode parafin dan pewarnaan HE. Hasilnya masih ditemukan semua tahapan perkembangan folikel (folikel primordial, primer, preantral, dan antral) pada semua kelompok perlakuan. Jumlah folikel pada 5, 7 atau 9 hari pascatransplantasi menurun nyata (p&lt;0,05) kecuali jumlah folikel primordial pada kelompok 5 hari pascatransplantasi (634,7±56,88) tidak berbeda nyata dengan kontrol (683,7±61,55). Dapat disimpulkan bahwa dinamika folikel ovarium domba pascatransplantasi pada kelinci pseudopregnansi masih dapat ditemukan pada semua kelompok perlakuan. THE FOLLICLE DYNAMICS OF EWE OVARIAN POST-INTRAUTERINE TRANSPLANTATION TO PSEUDOPREGNANCY RABBIT ABSTRACT The objective of this study was to evaluate the follicle dynamics of ewe ovarium post-intrauterine transplantation to pseudopregnanty rabbit. The experiment was concerned with the 1st or 7th days of pseudopregnancy to receive the ewe ovarium. After 5, 7, and 9 days transplantation the ewe ovarium were recollected. In order to determine the follicle dynamics of ewe ovari post-intrauterin transplantation and to count the number of each stage, the ewe ovari was prepared using the paraffin methods and staining with HE. The results showed all stages of the follicle dynamics (Primordial, Primary, Preantral and Antral follicle stages) were still found in all groups of treatment. The number of follicles decreased significantly (p&lt;0.05) except the number of Primordial follicles of the 5 days post transplantation (634.7±56.88) was not significantly different (p&lt;0.05) compared to the control group (683.7±61.55). It could be concluded that the follicle dynamics of ewe ovarium post-intrauterine transplantation in pseudopregnancy rabbit still found in all groups of the treatment.
VIABILITAS OOSIT DOMBA PASCATRANSPLANTASI OVARIUM DOMBA DALAM UTERUS KELINCI PSEUDOPREGNANT Sumarmin, Ramadhan; Winarto, Adi; Yusuf, Tutty Laswardi; Boediono, Arief
Majalah Ilmiah Peternakan Vol 11, No 1 (2008)
Publisher : Majalah Ilmiah Peternakan

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Abstract

ABSTRAK Telah dilakukan penelitian untuk mengetahui viabilitas oosit domba yang dikoleksi dari ovarium domba pascatransplantasi intrauterus pada kelinci pseudopregnant. Ovarium domba ditransplantasikan dalam uterus kelinci pseudopregnant pada hari pertama bunting semu dan kemudian diambil kembali setelah lima (P5) atau tujuh (P7) hari transplantasi. Sebagai kontrol digunakan ovarium segar. Oosit dikoleksi dari ovarium pascatransplantasi dengan metode slicing (pencacahan) di dalam media phosphate buffer saline (PBS) yang disuplementasi dengan 5% fetal bovine serum (FBS) dan 100 IU penicillin-streptomisin/ml. Oosit hasil koleksi selanjutnya dimaturasi dalam Tissue Culture Medium (TCM)-199 yang disuplementasi dengan 10% FBS, 10 IU follicle stimulating hormone (FSH)/ml dan 100 IU penicillin-streptomisin/ml. Oosit selanjutnya diinkubasi dalam inkubator CO2, dengan kandungan CO2 5% dan suhu 38ºC, selama 24 jam. Setelah dimaturasi, oosit diwarnai dengan aceto-orcein 2% untuk menentukan status inti oosit. Hasilnya memperlihatkan bahwa oosit yang mampu mencapai tahapan perkembangan dengan status inti M-II setelah maturasi pada P5 (35,05%) dan P7 (35,24%) nyata lebih sedikit (p&lt;0,05) dibandingkan dengan kontrol (56,65%). Dapat disimpulkan bahwa viabilitas oosit domba pascatransplantasi di dalam uterus kelinci pseudopregnant masih ditemukan meskipun persentasenya lebih rendah. EWE OOCYTE VIABILITY FROM EWE OVARIAN AFTER INTRAUTERINE TRANSPLANTATION TO PSEUDOPREGNANT RABBIT ABSTRACT The aim of the present study was to investigate the ewe oocyte viability from ewe ovary after intrauterine transplantation to pseudopregnant rabbit. The ewe ovary was transplanted intrauterine on day 1 to pseudopregnant rabbit and oocytes recollected on five (P5) or seven (P7) days posttransplantation. The fresh ovary was used as the control. The oocytes were collected from the ovaries by slicing method in Phosphate Buffer Saline (PBS) supplemented with 5% of Fetal Bovine Serum (FBS), and 100 IU/ml of penicillin-streptomycin. Oocytes were matured in Tissue Culture Medium (TCM)-199 supplemented with 10% FBS, 10 IU/ml of Follicle Stimulating Hormone (FSH), and 100 IU/ml of penicillin-streptomycin. Oocytes were incubated in CO2 incubator with 5% CO2, 38°C for 24 h. After maturation, the oocytes were stained with 2% aceto-orcein to determine the nuclear oocytes status. The result showed that the oocytes could reach the M-II phase from P5 (35.05%) and P7 (35.24%) decreased significantly (p&lt;0.05) compared to the control (56.65%). However it can be concluded that the oocytes viability still preserved intrauterine in pseudopregnant rabbit.
INJEKSI SPERMATOZOA DOMBA HASIL PENGERINGBEKUAN KE DALAM SEL TELUR MENGGUNAKAN TEKNIK INTRACYTOPLASMIC SPERM INJECTION (ICSI) INJECTION OF FREEZE-DRIED RAM SPERMATOZOA INTO OOCYTES USING INTRACYTOPLASMIC SPERM INJECTION, ICSI Saili, Takdir; Agus Setiadi, Mohamad; Agungpriyono, Srihadi; Boediono, Arief
Jurnal Veteriner Vol 8, No 1 (2007)
Publisher : Jurnal Veteriner

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Abstract

Pada penelitian ini dikaji kemampuan spermatozoa domba hasil pengeringbekuan untuk melakukan dekondensasi dan membentuk pronukleus setelah diinjeksikan ke dalam oosit dengan menggunakan teknik intracytoplasmic sperm injection (ICSI). Metode pewarnaan aceto lacmoid digunakan untuk mengevaluasi kejadian dekondensasi dan pembentukan pronukleus pada oosit setelah ICSI. Hasil penelitian menunjukkan bahwa spermatozoa hasil pengeringbekuan dapat melakukan dekondensasi (2%) dan mendukung pembentukan 1PN (34%) tetapi belum mampu mendukung pembentukan 2PN setelah diinjeksikan ke dalam oosit. Sedangkan injeksi dengan menggunakan spermatozoa segar mampu mendukung pembentukan 2PN (30%) dan 1PN (40%). Sebagai kesimpulan dapat dikemukakan bahwa spermatozoa hasil pengeringbekuan mampu melakukan dekondensasi dan mendukung pembentukan 1PN setelah ICSI
SEBARAN ANTIAGLUTININ SPERMATOZOA DALAM PLASMA YANG DIKOLEKSI DARI EPIDIDIMIS DAN EJAKULAT DOMBA THE DISTRIBUTION OF SPERM ANTIAGGLUTINI IN PLASMA COLLECTED FROM EPIDIDYMIS AND EJACULATE OF RAM Haviz, Muhamad; Agungpriyono, Srihadi; Boediono, Arief; Fahrudin, Mokhamad; Setiadi, M Agus
Jurnal Veteriner Vol 8, No 1 (2007)
Publisher : Jurnal Veteriner

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Abstract

Penelitian untuk mengetahui tingkat aglutinasi antarkepala spermatozoa dan sebaran antiaglutinin dalam plasma asal epididimi dan ejakulat telah dilakukan untuk mengoptimalkan pemanfaatannya dalam fertilisasi in vitro. Tingkat aglutinasi dan aktivitas antiaglutinin plasma epididimis dan ejakulat domba dihitung dengan cara menghitung jumlah aglutinasi antarkepala spermatozoa setelah diinkubasikan selama 1, 3, 5, dan 7 jam in vitro dalam media Krebs Ringer- HEPES.
PRODUCTION OF EMBBRYONIC STEM CELLS FROM INNER CELL MASS OF BLASTOCYST ISOLATED BY ENZYMATIC AND IMMUNOSURGERY METHODS Mata Hine, Thomas; Supriatna, Iman; Sajuthi, Dondin; Boediono, Arief
Jurnal Veteriner Vol 9, No 1 (2008)
Publisher : Jurnal Veteriner

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Abstract

The objective of the research is determining the ICM isolation method to produce ESC. Blastocyst stage of DDy mice embryos were used in this study. Zona pellucida of blastocysts were removed by 0.25% pronase, the ICM isolation were done by enzimatic or immunosurgery method, and then they were cultured in DMEM-high glucose supplemented with mercaptoethanol, gentamycin, fetal bovine serum, and cumulus cells as feeder layer. The result of the research indicated that immunosurgery method yielding attachment rate and number ESC colony 93.85% and 43.08%, respectively, higher (P&lt;0.05) than enzimatic method that weree 79.63% and 18.52%, respectively, but the viability of ICM cells were equal (P &gt;0.05) that are 93.59% in enzymatic method and 98.56% in immunosurgery method. This research concluded that immunosurgery more effective method for isolation of ICM and ESC production than enzymatic method.
Co-Authors . AULANI’AM . Herdis . Nurhidayat . WIDJIATI . Yulnawati Achmad Selamet Aku Adi Winarto Adrien Jems Akiles Unitly, Adrien Agus Setiadi Al Azhar AL-AZHAR AL-AZHAR Alfred O. M. Dima, Alfred O. M. Amrozi . Amrozi a Andriyanto A Aryani Sismin Satyaningtijas, Aryani Astini, Wining Aucky Hinting Bambang Kiranadi Bayu Rosadi Bibiana W Lay BIBIANA WIDIATI LAY Boenjamin Setiawan Budiariati, Vista Budiono, Dwi Caroline T. Sardjono Chairun Nisa Citra Noviana Dedy D. Solihin, Dedy D. DEDY DURYADI SOLIHIN Djaswadi Dasuki Djoko Walujo DONDIN SAJUTHI Dwi Haryadi Dwijayanti, Adisti EVY AYU ARIDA Fadjar Satrija Farid A. Moeloek Farizah, Nuril Ferry Sandra Fitrianto, Alif Iman Hadi, Restu S. Harry Murti HERA MAHESHWARI Herdis . herdis herdis Heri Sujoko Heru Setijanto I Ketut Mudite Adnyana I Ketut Mudite Adnyane I Ketut Suatha I Wayan Batan Ichsan Ichsan IMAN SUPRIATNA Indra Bachtiar Indra Kusuma Irma Suryani ITA DJUWITA Ita Fauzia Hanoum, Ita Fauzia Juliandi, Berry KARTINI ERIANI Kartiwa, R. Angga Ketut Adnyane Mudite KUSDIANTORO MOHAMAD LATIFAH KOSIM DARUSMAN Lea Tarliyah Lubis, Alkaustariyah Lubis, Andri Maruli Tua M Agus Setiadi M Agus Setiadi Maman Surachman Min Rahminiwati Miraprahesti, Retti N. Mohamad Agus Setiadi Mohamad Agus Setiadi Mohamad Fakhrudin Mohammad Ghozali, Mohammad Mokhamad Fahrudin MOZES R. TOELIHERE Muhamad Haviz MUHAMMAD AGUS SUPRAYUDI Muhammad Gunawan Muhammad Haviz MUHAMMAD RIZAL MULYOTO PANGESTU NASTITI KUSUMORINI Nining Handayani Nining Handhayani Nurhidayat Said Nursanti, Risa Rachmat Herman Rakhmawati, Handina Ramadhan Sumarmin Rangga Setiawan Ratih Rinendyaputri Ridi Arif Ridlo, Muhammad Rosyid Rimayanti - Rini Widyastuti, Rini Riris L. Puspitasari Ronny Rachman Noor Satya Gunawan Shofwal Widad Situmeang, Adrian SONY HERU SUMARSONO Sri Catur Setyawatiningsih Srihadi Agungpriyono Supar - Supar . Sutarya Enus SUTIMAN BAMBANG SUMITRO Suyatna, Frans Dhyanagiri Syamsunarno, Mas Rizky Anggun Adipurna TAKDIR SAILI Taufik Jamaan Thomas Mata Hine Thomas Mata Hine Trevino A. Pakasi TRINIL SUSILOWATI Tutik Wrediati TUTIK WRESDIYATI Tutty Laswardi Yusuf Tutty Laswardi Yusuf TUTY LASWARDI YUSUF Utami, Adkhilni Vincentia Maria Wahono Esthi Prasetyaningtyas Wahono Esti Prasetyaningtyas Wahono Esti PrasetyoningtyaserB Wasmen Manalu Widjiati - Yaprianto, Kelvin Yuhara Sukra Yulnawati . YULNAWATI YULNAWATI Zairin JR, Muhammad