Studies On HALR Transfecting Sheep Embryonic Fibroblasts Cell Mediated By Liposome

In our experiments, we studied how the factors affected the transfaction efficiency and what effects the liposome method took on sheep embryonic fibroblasts cells in order to increase transfaction efficiency and grand for sheep biology reactor.1. In vitro culture system of sheep embryonic fibroblasts (sFFCs) and preferential.transfection method was investigated in this study.cell line of sheep embryonic.fibroblasts (sFFCs) was established by embryo of 30th sheep successfully. Moreover, there was 46% of cells transfected positively by use of optimum condition of transfection. According to the behest of cell line,the present study performed a biological study, the results as follows: character of classic frbroblast, and could be purified for their diferent sensibility to trypsin.2. The growth curve of sFFCs show the shape of"s"and went through lag phase, log phase, plateau phase.3. sFFCs were frozen in DMEM with serum and adding 10% DMSO, The survival rates of sFFCs well, they are both can meet the needs of frbroblast passage and log-term preservation.4. Karyotype analysis indicated that sFFCs had 2n=54 chromesomes, this culture system was suitable and safe to sFFCs, and keep relatively stable karyotype.5. pNIGBA could efficiently transfected the sFFCs by LipofectamineTM Reagent. The present study optimize several factors influence transfection efficiency; confluence rate of sFFCs (30%-50%), proportion of plasmid DNA (1.0μg) /liposome (3.0μ1), concentration of G418 (800μg/ml), and transfection eficiency can gradually increased to 46%.