The Orientation Of The Mouse Es Cells Induced To Differentiate With The Department Of Experimental Construction

In these years, because of its totipotency, manipulation and plastics, the embryonic stem (ES) cells have been the focus of researches in the fields of medical and life science. With the help of ES cells, scientists are trying to set up the cell and animal models of diseases such as diabetes, Alzheimer's disease and Parkinson's disease, produce the transgenic animals. As for these researches, the most important thing that must be resolved is the resource of ES cells and the differentiation of ES cells induced to specific cell. In order to do such research, we must have sufficient ES cells; especially for the production of transgenic animals, ES cells within 10 passages are required. And if for the study of cell therapy, the problem of other differentiated cells coexisting in the wanted cells-derived from ES cells must be resolved.In our study, ES-D3 cells were used and proliferated by cell culture, and then we induced the ES cells to differentiate into the neuron-like cells by two methods. First, we tried to explore the effects of embryonic bodies (EBs) on the differentiation of ES cells, further the intrinsic differentiation mechanism of ES cells. The EBs of different days gained by means of hanging, suspending and suspending following hanging were processed by all-trans retinoic acid (RA) for 4 days. During this period, we observed the differentiation process and compared the differentiation ratio of ES cells processed by means of hanging, suspending for several days. Based on this study, we induced the EBs by RA of different concentration in order to study the effect of RA on the differentiation of ES cells into neuron-like cells. The results suggested that if the EBs was induced by means of hanging for 3d and suspending for Id by RA in the concentration of 10~6M, a comparatively higher differentiation ratio will be got.In order to supply the ES cells needed for the research of transgenic animals in our labortaory in future, we did some work of isolating ES cells from mouse. We used the ICR and Kunming mice and got the embryos of 3.5dpc by means of superovulation, then cultured the embryos on the feeder layer derived from ICR or Kunming mice, dispersed the inner cell masses(ICMs) or ES cell colony in theappropriate time. In this period, we analized the effects of feeder layer, trypsin and embryos isolated from uteri of different varietal mice on the ES cell lines. The results showed that the feeder layers prepared by ICR and Kunming mice had no significant effect on adherence of embryos and proliferation of ICMs. The trypsin, which affected the isolation of ES cells remarkedly, should be used with concentration of 0.125% trypsin+0.02% EDTA for KMSmin. Furthermore, it is more easy to isolate embryos and get the ES cells from ICR mouse than Kunming mouse.