Mokhamad Fahrudin
Departemen Anatomi Fisiologi Dan Farmakologi, Fakultas Kedokteran Hewan, Institut Pertanian Bogor, Jln Agathis, Dramaga, Bogor, Jawa Barat, Indonesia, 16680

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Journal : Jurnal Veteriner

Perkembangan Praimplantasi Embrio Mencit dengan Materi Genetik yang Berasal dari Parental, Maternal, dan Inti Sel Somatik (PRE-IMPLANTATION DEVELOPMENT OF MOUSE EMBRYO WITH GENETIC MATERIAL DERIVED FROM PARENTAL, MATERNAL AND SOMATIC CELL NUCLEUS) Harry Murti; Mokhamad Fahrudin; Mohamad Agus Setiadi; Boenjamin Setiawan; Arief Boediono
Jurnal Veteriner Vol 15 No 1 (2014)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Cloned embryo and parthenogenetic embryo are a potential source of stem cells for regenerativemedicine. Stem cells derived from those embryos are expected to overcome the ethical issues to the use offertilization embryos for therapeutic purposes. The pre-implantation development is a critical step fordeveloping embryos reach the blastocyst stage. The objectives in vivo of this research are to produce mousecloned embryo, parthenogenetic embryo, and fertilized embryo and to study stages of  in vitro pre-implantation development culture. In vivo fertilized embryos, mouse oocytes, and cumulus cells were usedin this study. Treatment was performed on female mice superovulated with PMSG and hCG injections.Two-cell stage of in vivo fertilized embryos were collected on the second day post hCG injection. Clonedembryos were produced through Somatic Cell Nuclear Transfer (SCNT), which included enucleation, nucleartransfer and artificial activation. Parthenogenetic embryos were produced with artificial activationtechnique. The result of the research indicated that SCNT application was able to produce cloned embryos which could develop to blastocyst stage (3,2%). In addition, artificial activation of oocytes could produceparthenogenetic embryos which were able to develop up to the blastocyst stage (8,6%). In conclusion,efficiency level of parthenogenetic embryos that is able to reach the blastocyst stage was higher than in thecloned embryos. Fertilized embryos shows a better development and more efficient compared to in vitrocloned embryos and parthenogenetic embryos cultures.
STUDY ON SPERM AGGLUTINATION WITH CHARACTERIZATION OF PLASMACOLLECTED FROM EPIDIDYMIS AND EJACULATE IN RAM Muhammad Haviz; Arief Boediono; M Agus Setiadi; Srihadi Agungpriyono; Mokhamad Fahrudin
Jurnal Veteriner Vol 9 No 4 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

This study was designed to optimalize the use of epididymal or ejaculate sperm and plasma for in vitro fertilization, that sperm agglutination was found at preparation. The rate of sperm agglutination was calculated the head-to-head sperm agglutination that were incubated in Krebs Ringer-(N-(2hydroxyethyl)piperazine-N’-(2-ethenesulfonic acid) or KR-HEPES medium in 38.50C with 5% CO2 at 1, 3, 5 and 7 hours culture in vitro. The rate of head-to-head sperm agglutination were decreased with time treatments. The cauda of sperm agglutination was lower than that caput, corpus epididymal and ejaculate sperm with statistically significant (P<0.01). These result reflected that distribution of anti-agglutinin might be higher in cauda epididymal than that other areas. Number of protein were characterize with SDS-PAGE as follow 11 bands in caput epididymal, 9 bands in corpus epididymal, 2 bands in cauda epididymal and 4 bands in seminal plasma. The higher distribution of protein was found at range 25-40 kDa in epididymal plasma of ram. However, further investigation should be conducted to determine presumptive anti-agglutinin by advance method.
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 Muhamad Haviz; Srihadi Agungpriyono; Arief Boediono; Mokhamad Fahrudin; M Agus Setiadi
Jurnal Veteriner Vol 8 No 1 (2007)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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.
Tingkat Proliferasi Primordial Germ Cells secara In Vitro dalam Medium Kultur dengan Penambahan Leukemia Inhibitory Factor (IN VITRO PROLIFERATION RATE OF MICE PRIMORDIAL GERM CELLS IN THE CULTURE MEDIUM WITH ADDITION OF LEUKEMIA INHIBITORY FACTOR) Wahono Esthi Prasetyaningtyas; Ni Wayan Kurniani Karja; Srihadi Agungpriyono; Mokhamad Fahrudin
Jurnal Veteriner Vol 20 No 4 (2019)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (136.304 KB) | DOI: 10.19087/jveteriner.2019.20.4.526

Abstract

Primordial germ cells (PGCs) are precursors for gamete cells. The totipotency of PGCs allows them to be used as a model for studying cancer and infertility. The study aimed to examine the characteristics of the mice fetus as a source of PGCs, proliferation rate of PGC and the role of LIF in vitro culture of PGCs. This study used genital ridges from 26 fetuses at 13.5 days post-coital (dpc) to isolate the PGCs. Genital ridges dissociation using 0.1% of trypsin and in vitro culture was carried out using the Dulbecco Modified Eagle Medium (DMEM) and incubated at 37 0C and 5% CO2 atmosphere. The fetus was measured and weighed to determine the normal development of the fetus and continued with the identification of the genital ridges after laparotomy performed under a stereomicroscope. Proliferation rate was measured by calculating Population Doubling Time (PDT), and cell viability was observed after in vitro culture for six days. The effect of adding 1000 IU/ml of leukemia inhibitory factor (LIF) was evaluated from two types of treatment in the medium, 1) DMEM added with 15% of fetal calf serum (FCS) (DMEM + S15%) and 2), DMEM was supplemented with 15% of FCS and 1000 IU/ml LIF (DMEM + S15% + LIF1000 IU/ml). Immunohistochemistry staining was carried out on the third-, sixth- and ninth-day of culture to detect the expression of Oct-4 in the PGCs, then cells were counted. The results showed that the fetus as a source of PGCs had normal development. The fetal sizes were 11 mm, and male and female genital ridges could be distinguished by morphology at the age of 13.5 dpc. The proliferation of PGCs was relatively slow with a 1.3 day PDT value with the viability of around 85%. Culture of PGCs with DMEM + S15% treatment showed the percentage of PGCs that expressing Oct-4 decreased from the third day of culture to the ninth day of culture. The culture of PGCs in DMEM + S15% + LIF 1000 IU / ml treatment showed that the percentage of PGCs that expressed Oct 4 increased on the sixth day of culture and decreased on the ninth day of culture. It can be concluded that the addition of LIF can maintain the number of PGCs until the sixth day of culture. LIF is thought to play a role in the regulation of proliferation of PGCs through receptors of LIF (RLIF) and glicoprotein (gp) 130 receptors.
Mice Oocytes Respond after Vitrification Followed by Artifical Activation Using a Various Concentration of Strontium Chloride and Cytochalasin B Muhammad Rosyid Ridlo; Rini Widyastuti; Alkaustariyah Lubis; Mokhamad Fahrudin; Arief Boediono
Jurnal Veteriner Vol 19 No 3 (2018)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Oocyte vitrification is the important part of gamete preservation for further purpose. The objective of this study was to evaluate the response or development of vitrified-mice oocyte following activation using various concentrations of Strontium Chloride (Sr Cl2). Oocytes were collected from superovulation-induced female mice. Oocytes vitrification was then performed using a gradual equilibration of 2 M Ethylene Glycol in 0.25 M sucrose and 7 M Ethylene Glycol on 0.5 sucrose. Subsequently, the vitrified oocytes were thawed and activated using various Strontium Chloride concentration in each group. Control 1 is unvitrified oocyte and without Sr Cl2. Control 2 is unvitrified oocyte then activated by 20 mM Sr Cl2. Zero (0) mM Sr Cl2 is vitrified oocyte without Sr Cl2. Group Ten (10) mM is vitrified oocyte then activated by 10 mM Sr Cl2. Group Twenty (20) mM is vitrified oocyte then activated by 20 mM Sr Cl2. The viability of vitrified-thawed oocytes was observed based on ooplasm integrity. Whereas the oocytes respond to artificial activation was observed based on pronucleus formation after 10 hours of activation. The result showed that 39% of oocyte degenerated following vitrification. The respond of vitrified-thawed oocytes following artificial activation using Strontium Chloride was significantly lower compared to fresh oocytes (p<0,05). Interestingly the highest percentage of activated oocytes (36.36%) was present in a group achieved 20 mM Strontium Chloride. As conclusion is Strontium Chloride 20mM has a best result (36,36%) to activate vitrified oocyte than 0 mM and 10 mM of Strontium Chloride.
Perbedaan Morfologi dan Ekspresi Dazl dan Vasa pada Sel Germinal Fetus dan Anak Mencit Jantan Wahono Esthi Prasetyaningtyas; Ni Wayan Kurniani Karja; Mokhamad Fahrudin; Kusdiantoro Mohamad; Srihadi Agungpriyono
Jurnal Veteriner Vol 24 No 1 (2023)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19087/jveteriner.2023.24.1.49

Abstract

Sel germinal merupakan salah satu sumber sel yang masih bersifat totipotensi dan berperan dalam pembentukan organisme baru. Morfologi dan ekspresi protein pada sel germinal bersifat dinamis bergantung pada umur dan tahap perkembangan. Penelitian ini bertujuan untuk mengamati perubahan morfologi dan ekspresi protein sebagai marka sel germinal jantan pada fetus umur 13,5 hari pascakawin (days post coital/ dpc) dan anak mencit umur lima hari pascalahir. Hasil Rigi kelamin dan testis diisolasi dari mencit umur 13,5 dpc dan 5 hari. Jaringan kemudian dipreparasi histologi rutin, dan diwarnai dengan pewarnaan hematoksilineosin (HE), sedangkan untuk mengidentifikasi keberadaan protein Dazl, Vasa dan Oct4, jaringan diwarnai dengan pewarnaan imunohistokimia menunjukkan morfologi sel germinal jantan pada fetus mencit umur 13,5 dpc dan anak mencit umur lima hari pascalahir sama-sama berbentuk bulat oval. Namun, sel germinal jantan pada mencit umur lima hari pascalahir berukuran lebih besar, jumlah yang lebih sedikit dan terletak jauh dari membran basal. Pada sel germinal jantan umur 13,5 dpc menunjukkan positif lemah terhadap antibodi Oct 4 dan DAZL serta positif kuat terhadap antibodi Vasa. Pada umur lima hari, sel germinal jantan menunjukkan positif kuat terhadap antibodi Oct-4 dan DAZL, serta positif lemah terhadap antibodi Vasa. Simpulan dari penelitian ini adalah morfologi dan ekspresi marka sel germinal dipengaruhi oleh tahapan pertumbuhan dan perkembangan sel-sel germinal. Vasa dapat digunakan sebagai marka untuk sel germinal umur 13,5 dpc dan DAZL sebagai marka untuk sel germinal umur lima hari pascalahir.