Abstract
Cytoplasm injection cloning technology (CICT) is an efficient technique for evaluating the developmental potential of cloned embryos. In this study, we investigated the effects of donor cell type on the developmental potential and quality of cloned bovine embryos. Adult fibroblasts (AFs) and embryonic cells (ECs) were used as donor cells to clone bovine embryos using CICT. We initially used AF cells to develop cloned embryos and then cultured the cloned day-8 blastocysts for 10 days to obtain ECs as donor cells for second embryo cloning. We found that the bovine blastocysts cloned using AF cells had significantly reduced developmental rates, embryo quality, and ratios of inner cell mass (ICM) to the total number of cells compared to those using ECs as donor cells. Furthermore, there were significant differences in the DNA methyltransferase-, histone deacetylation-, apoptosis-, and development-related genes at the blastocyst stage in embryos cloned from AFs compared to those in embryos cloned from ECs. Our results suggest that using ECs as donor cells for nuclear transfer enhances the quantity and quality of cloned embryos. However, further investigation is required in terms of determining pregnancy rates and developing cloned embryos from different donor cell types.
Highlights
Cloning refers to the processes used to produce the exact genetic replica of a biological entity
Cytoplasm injection cloning technology (CICT) is a type of Somatic cell nuclear transfer (SCNT), where extra cytoplasm is injected into the enucleated oocyte with the donor cell, which improves the quality of cloned embryos [7]
We identified that H3K9me2 expression in Adult fibroblasts (AFs) was significantly higher than that in the embryonic cells (ECs), which indicates transcriptional repression in the AFs (Figure 1A)
Summary
Cloning refers to the processes used to produce the exact genetic replica of a biological entity. Considerable differences have been found in the ability of different cell lines and cell types to be reprogrammed and in the developmental competence of cloned embryos produced from them. It is still unclear which cell type is the most efficient for SCNT. We used adult fibroblasts (AFs) and embryonic cells (ECs) as donor cells to analyze the quality of cloned bovine embryos developed using CICT. In the present study, we compared the effectiveness of donor cell types on the developmental potential and blastocyst quality of cloned bovine embryos using the CICT
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