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        The Effects of Donor Cell Type and Culture Medium on in vitro Development of Domestic Cat Embryos Reconstructed by Nuclear Transplantation

        Fahrudin, Mokhamad,Otoi, Takeshige,Karja, Ni Wayan Kurniani,Murakami, Masako,Suzuki, Tatsuyuki Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.8

        In this study we explored the possibility of performing nuclear transfer in the domestic cat and assessed the ability of different culture media to support in vitro development of reconstructed cat embryos. Donor somatic cells were derived from cultured cumulus cells or explants of oviduct tissue, and recipient cytoplasts from in vitro matured oocytes. A higher percentage of cleavage (84.6% and 86.5%) and development to the morula stage (35.9% and 44.2%) was found when reconstructed embryos receiving cumulus or oviduct cells were cultured in MK1 medium, compared with those cultured in CR1aa (58.7% and 72.5%, 13.8% and 13.6%, respectively). There was no significant difference between MK1 and CR1aa media with respect to the proportion developing to the blastocyst stage (15.4% and 17.3% vs 6.8% and 8.6%, respectively, p>0.05). There was no significant effect (p>0.05) of donor cell type (cumulus and oviduct cells) on the rates of fusion (65.0% and 52.5%), cleavage (84.6% and 86.5%), development to the morula (35.9% and 44.2%), and blastocyst (15.4% and 17.3%) stages when reconstructed embryos were cultured in MK1 medium. Similar results were found for the reconstructed embryos cultured in CR1aa medium. These results show that culture medium has a significant impact on the early development of reconstructed cat embryos, whereas donor cell type does not have a significant effect.

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        Conditioned medium of E17 rat brain cells induced differentiation of primary colony of mice blastocyst into neuron-like cells

        Vista Budiariati,Ratih Rinendyaputri,Ariyani Noviantari,Noer Muhammad Dliyaul Haq,Dwi Budiono,Diah Nugrahani Pristihadi,Berry Juliandi,Mokhamad Fahrudin,Arief Boediono 대한수의학회 2021 Journal of Veterinary Science Vol.22 No.6

        Background: Conditioned medium is the medium obtained from certain cultured cells and contained secretome from the cells. The secretome, which can be in the form of growth factors, cytokines, exosomes, or other proteins secreted by the cells, can induce the differentiation of cells that still have pluripotent or multipotent properties. Objectives: This study examined the effects of conditioned medium derived from E17 rat brain cells on cells with pluripotent properties. Methods: The conditioned medium used in this study originated from E17 rat brain cells. The CM was used to induce the differentiation of primary colonies of mice blastocysts. Primary colonies were stained with alkaline phosphatase to analyze the pluripotency. The morphological changes in the colonies were examined, and the colonies were stained with GFAP and Neu-N markers on days two and seven after adding the conditioned medium. Results: The conditioned medium could differentiate the primary colony, beginning with the formation of embryoid-body-like structure; round GFAP positive cells were identified. Finally, neuron-like cells testing positive for Neu-N were observed on the seventh day after adding the conditioned medium. Conclusions: Conditioned medium from different species, in this case, E17 rat brain cells, induced and promoted the differentiation of the primary colony from mice blastocysts into neuron-like cells. The addition of CM mediated neurite growth in the differentiation process.

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