Reproduction Abstracts

Introduction: Nuclear transfer (NT) has used for generating cloned animals or genetically modified animals. However, the efficiency has remained low, because of epigenetic errors that occur during donor cell reprogramming. Mesenchymal stem cells (MSCs) were known as undifferentiated state cell compared to somatic cells. Thus MSCs can reduce the chance of error that can be occurred during reprogramming process and can be easily isolated from adult, while embryonic stem cells are not. For these reasons, undifferentiated stem cells may be more suitable as donor cells for NT than fully differentiated somatic cells. To increase the rates for cloned animal production, MSCs will be a good donor cell source for successive NT.

Materials and methods: In this study, we isolated MSCs from porcine fetal femur and explants on the plate directly. Putative femur-derived MSCs were first observed after 5–6 days of culture. Cells at passage 3–5 were characterized by RT-PCR for marker gene expression and differentiation capacity of osteogenesis and adipogenesis and then those cells were used as a donor cells.

Results and discussion: Femur-derived MSCs showed fibroblast-like morphology. Positive markers for MSC (CD29, CD44, CD90, CD105, CD144) was highly expressed in this cell, whereas negative markers (CD14, CD34, CD45) were not detectable. Especially, expression of major MSC surface molecules (CD90, CD105) were observed by FACS. According to the osteogenesis and adipogenesis from MSCs, differentiation potential was verified. These results suggest that MSCs from porcine fetal femur were successfully isolated, and might be a good donor cells for producing porcine NT embryo.