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 : HAYATI Journal of Biosciences

Production and Characterization of Mouse Diploid Parthenogenetic Blastocyst Developed in Phosphate-Free Medium Budiariati, Vista; Budiono, Dwi; Fahrudin, Mokhamad; Juliandi, Berry; Rinendyaputri, Ratih; Boediono, Arief
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (396.143 KB) | DOI: 10.4308/hjb.27.2.89

Abstract

Parthenogenesis is an artificial oocytes activation process without paternal contribution. Blastocyst, derived from parthenogenesis, is one of potential source for pluripotent stem cell propagation. Unfortunately, previous studies reported that parthenogenetic embryo did not achieve exhilarating blastocyst rate. One of the component that predicted inhibit parthenogenetic embryo development is phosphate. Therefore, we try to modify culture medium in order to overcome that problem. The aim of this research was to produce and analyze the characteristics of parthenogenetic blastocyst developed in phosphate-free medium. Mouse oocytes obtained from adult female DDY by superovulation. The activator was strontium chloride 10 mM and diploidization with cytochalasin B 5 μg/ml. Medium for activation and culture medium were modified rat 1 cell embryo medium (MR1ECM) which is phosphate free. The results showed that parthenotes that were cultured in phosphate free medium reached higher blastocyst rate compared to the other groups. The increase of phosphate in culture medium lead to impaired parthenogenetic embryos development. Further experiment was made to analyze the differences between fertilized and parthenogenetic embryo in this medium. The experiment showed that diploid parthenogenetic could achieve high blastocyst rate (30.9±1.3%). The quality of diploid parthenogenetic blastocyst, based on cells number, viability, and ICM ratio, was lower than fertilized blastocyst.
Inhibitory Effect of Iodoacetate on Developmental Competence of Porcine Early Stage Embryos In Vitro NI WAYAN KURNIANI KARJA; MOKHAMAD FAHRUDIN; KAZUHIRO KIKUCHI
HAYATI Journal of Biosciences Vol. 16 No. 1 (2009): March 2009
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (61.946 KB) | DOI: 10.4308/hjb.16.1.25

Abstract

In porcine preimplantation embryos, glucose-utilizing pathways are reported active throughout their development. The aim of this study was to test the involvement of glycolysis activity in early stage of porcine embryo development by the addition of iodoacetate, a glycolytic inhibitor, to culture medium. In vitro matured and fertilized oocytes were cultured for the first 2 days in iodoacetate at concentrations of 0, 1, or 2.5 µM. The proportion of viable embryos on day 4 decreased when Ia was added at concentration of 2.5 µM (P < 0.05). The development of cleaved embryos to the blastocyst stage on day 6 was significantly reduced by 1 µM iodoacetate and none of cleaved embryos in Ia-2.5 group develop to the blasyocyst stage (P < 0.05). More than 60% of embryos in Ia-2.5 group were arrested at the two-four-cell stage, and then arrested at the morula stage. These results indicate that glucose plays important roles in supporting the development of early stage of porcine embryos and that the inhibition of glucose metabolism may disrupt the management of energy production, leaving the embryos incompetence to develop. Key words: iodoacetate, glucose, early stage embryos, in vitro culture
Production and Characterization of Mouse Diploid Parthenogenetic Blastocyst Developed in Phosphate-Free Medium Vista Budiariati; Dwi Budiono; Mokhamad Fahrudin; Berry Juliandi; Ratih Rinendyaputri; Arief Boediono
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (396.143 KB) | DOI: 10.4308/hjb.27.2.89

Abstract

Parthenogenesis is an artificial oocytes activation process without paternal contribution. Blastocyst, derived from parthenogenesis, is one of potential source for pluripotent stem cell propagation. Unfortunately, previous studies reported that parthenogenetic embryo did not achieve exhilarating blastocyst rate. One of the component that predicted inhibit parthenogenetic embryo development is phosphate. Therefore, we try to modify culture medium in order to overcome that problem. The aim of this research was to produce and analyze the characteristics of parthenogenetic blastocyst developed in phosphate-free medium. Mouse oocytes obtained from adult female DDY by superovulation. The activator was strontium chloride 10 mM and diploidization with cytochalasin B 5 μg/ml. Medium for activation and culture medium were modified rat 1 cell embryo medium (MR1ECM) which is phosphate free. The results showed that parthenotes that were cultured in phosphate free medium reached higher blastocyst rate compared to the other groups. The increase of phosphate in culture medium lead to impaired parthenogenetic embryos development. Further experiment was made to analyze the differences between fertilized and parthenogenetic embryo in this medium. The experiment showed that diploid parthenogenetic could achieve high blastocyst rate (30.9±1.3%). The quality of diploid parthenogenetic blastocyst, based on cells number, viability, and ICM ratio, was lower than fertilized blastocyst.
Interspecies Reconstructed Embryonic Cell Interaction between Campbell Hamster (Phodopus campbelli) and Mice (Mus musculus) Noer Muhammad Dliyaul Haq; Diah Pristihadi; Vista Budiariati; Dwi Budiono; Al Mukhlas Fikri; Mokhamad Fahrudin; Cece Sumantri; Arief Boediono
HAYATI Journal of Biosciences Vol. 28 No. 3 (2021): July 2021
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.28.3.185

Abstract

Interspecies embryo transfer is a seldom-used method to increase the successful conservation of endangered species. The study aimed to determine the potential development of interspecies reconstructed embryos. The present study used two animal models, Campbell hamsters (Phodopus campbelli) and mice (Mus musculus). The isolated inner cell mass (ICM) of hamster embryos were injected into the mice embryos. The embryos were transferred to the pseudopregnant mice using non-surgery embryos transfer methods. The fetuses were collected at day 13.5 of gestation for morphometric measurement and cytochrome b (Cyt b) analysis which used to determine the species of obtained fetuses. The results showed that the viability, pregnancy rate, and embryonic implantation ability of the interpecies reconstructed embryos did not differ significantly (p>0.05) compared to non-reconstructed embryos. Morphometric measurement showed that the crown-rump (CR) and the weight of fetuses in the reconstructed group were significantly higher than non-reconstructed group (p<0.05). According to Cytb analysis, the species of obtained fetuses were mice, while the population of hamster cells were found only in the blighted ovum (resorption). Therefore, it can be concluded that interspecies reconstructed embryos are able to implant. However, the population of mice cells are only found to develop.