| The application of the potential for ESC has been approved by the medicine, the field of basic research and the market of commercialization, because that the ESC has the multipotency and its self renewal. The study based on the latest progress of ESC and on our lab works on establishing oESC-like lines. In this paper, We investigated the rate of oESC-like lines with different developmental stages of blastocysts. And during the following process of culturing oESC-like, we added the LIF and bFGF in the culture media of oESC-like to observe the influences on proliferation, apoptosis of oESC-like and expression of multipotent candidate genes. The main research results as follows:We have isolated the in vitro culture blastocysts at different developmental stages (early blastocysts, extended blastocysts, hatached blastocysts) by the same method, The formation rates of primary oESC-like are 19.6%(3/12) with early blastocysts,37.5%(27/72) with extended blastocysts, and 25%(3/12) with hatached blastocysts (P<0.01).The assays of alkaline phosphatase are all positive in these experiments.We have added LIF and bFGF cytokines during culture oESC-like showed that the speed of proliferation in group bFGF was remarkbable compared with other experimental groups by the detection of proliferation and apoptosis of growing oESC-like for five days (P<0.01). while the proliferation of mLIF, hLIF and control group (N2B27) are not significant (P>0.05). We also found that LIF and bFGF had no influence in apoptosis (P>0.05). We detected the expression of multipotent genes by Real-time PCR and found that LIF could upregulate the expression of oESC-like multipotent candidate gene SOX2 (P<0.01), and downregulate the expression of OCT4(P<0.01). There are no diffirences to regulate the expression of multipotent candidate gene SOX2 and OCT4 between mLIF and hLIF cytokines (P>0.05). However, | bFGF could upregulate the expression of multipotent candidate genes, SOX2 and OCT4, in oESC-like lines (P<0.01). |
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