External pH is crucial in preserving sperm viability and ensuring fertilization during in vitro conditions. The purpose of this study was to determine the maximum pH value that can be tolerated by frozen-thawed Pasundan bull sperms and the effect on sperm quality. Around 250x10 6 sperms/mL of frozen-thawed Pasundan bull sperms were divided into ten equal aliquots, and each was diluted in the medium within a particular pH value. HCL or NaOH was added to the buffer media to create ten different solutions with varying pH values of 3, 4, 5, 6 as acidic, 7.2-7.4 as a control, and 8, 9, 10, 11, and 12 as alkaline. Furthermore, the samples were incubated for 5 minutes at room temperature within a particular pH medium before being immediately supplemented with a buffered medium to achieve a pH of 7.2-7.4. After 10 minutes of incubation at room temperature, all parameters were assessed. The results showed that sperm motility, viability, normal morphology, and acrosome intactness in sperms incubated in the acidic or alkaline media were significantly lower compared to control (p<0.05, respectively). Interestingly, the sperm still had a good tolerance to pHs 6 and 8. This tolerance was evidenced by all the parameters of sperms that were not sharply decreased compared to the control group. The significant loss of motility occurred at pHs 3 and 12. It could be concluded that frozen-thawed Pasundan bull sperms are still tolerable in pHs 4-11, but the sperm quality degrades as the acidity or alkaline level increases.

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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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.

Pasundan cattle are native Indonesian cattle located in West Java, Indonesia. These cattle have a superior trait in reproductive performance although reared in extreme environmental conditions with low-quality of feed, and resistance to parasite diseases than other cattle breeds. However, the ability to maintain Pasundan cattle genetic diversity, avoiding inbreeding, keeping the population healthy, and high reproduction efficiency are among the significant challenges currently occurred. The application of assisted reproductive technology (ART) seems promising to overcome those challenges and would impact the efficiency of genetic diversity conservation and preservation. This review aims to present the possibility to use the available ART in Pasundan cattle. The ART reviewed are the advanced technology of sperm cryopreservation, sex selection, artificial insemination, and in vitro embryo production. The sperm cryopreservation combined with sperm sexing is the most promising technology to maximize the use of genetic material of Pasundan cattle. Moreover, sexed sperm would have a promising impact on artificial insemination and/or embryo production to increase the cattle population. Furthermore, to protect genetic diversity, it is possible to apply the artificial reproductive technique and sperm or embryos conservation for genetic utilization in the future.