| Genetic background can affect development of animal embryos and development status of animals after birth. Metaphase ? oocytes collected from female mice can be artificially activated by chemical methods, then develop to blastocysts in vitro. The harvested blastocysts were cultured with ES cell medium and finally isolate pESCs. The genetic background can affect development of parthenogenetic embryos and isolation efficiency of pESCs. Previous studies have shown that genetic background can affect efficiency of pESCs in mice, but if it can affect the differentiation potential of pESCs is still unknown. In this study, we have detected gene expression of pluripotency and mitochondria of pESCs to explore the influence of genetic backgrounds on the quality of oocytes, at the same time, the results can provide references for further research about pESCs.(1) In this study, pESCs were established from oocytes of hybrid offspring of Kunming (KM 1/2) and 129/Sv mice, Kunming and C3H mice were used as object, and J1 ESCs and control. We first confirmed that J1 ESCs and pESCs have similar basic properties for morphology, cell totipotency of PCR and qPCR,immunofluorescence, differentiation ability in vitro. However there were significant differences in the expression levels of some pluripotent genes. The results showed that Expression level of Oct-4 in J1 ESCs was higher than that of KM×C3H F1 pESCs (p<0.05), Expression level of Sox2, H19, IGF2, KIF4, C-myc were significantly higher (p<0.01); Expression level of Sox2 and Nanog were higher in KM×129/Sv F1 pESCs, C-myc was significantly higher (p<0.01); In addition, the diameter of embryonic bodies (EBs) that formed jn KM×C3H F1 pESCs was significantly lower (p<0.05) and in KM×129/Sv F1 pESCs was significantly lower(p<0.01). Overall the expression levels of pluripotent genes in pESCs were significantly higher than that of J1ESC. This result demonstrated that the paternal genetic backgrounds affect the pluripotency of pESCs.(2) Detecting the proliferation ability of J1ESCs and pESCs in one week, at the same time, the gene expression level of Pnptl, Cox2, tim23, tom40,12sRNA and CytoC related to mitochondrial functions were detected by PCR and qPCR methods. The results showed that: Proliferation rate of KM×129/Sv F1 pESCs was significant lower than J1 ESCs in 2-5 days (p<0.05), in the 6th day the proliferation speed suddenly accelerated (P<0.05),however, KM×C3H F1 pESCs proliferation ability was significantly lower than that of J1 ESCs on daysl, 3, and 6 (P< 0.05), in the 4th and 5th days were extremely significantly lower (P<0.01), and on the seventh day the number of viable cells were decreased obviously of J1 ESCs and pESCs; Expression level of tim23 in J1ESCS was higher than that of KM×C3H F1 pESCs (p<0.05) Expression level of tom40 was significantly higher in KM×129/Sv F1 pESCs (p<0.05), contrary to which, Cyto C was significantly lower (p<0.01) and 12sRNA was significantly higher (p<0.01). Cell proliferation assay showed that the proliferation ability of J1 ESCs was faster than that of pESCs. The results discovered that paternal genetic background afect the function of Mitochondria. |
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