Title: Optimization of parameters of exogenous gene mediated by liposome to transfect yak mammary epithelial cells in vitro.
Abstract: [Objective] The aim of this study was to optimize conditions of exogenous gene mediated by liposome to transfect yak mammary epithelial cells in Vitro.[Method] Yak mammary epithelial cells were isolated and cultivated in Vitro by the methods of collagenase digestion and tissue adhesion.The expression of cytokeratin in yak mammary epithelial cell was detected by immunocytochemistry technique.With green fluorescence protein as the report gene,yak mammary epithelial cells were transfected with exogenous gene mediated by liposome,and the transfection efficiency in different cell fusion degree,liposome concentration,liposome/plasmid ratio and transfection time was statistically obtained to find the optimal transfection conditions.[Result] The results showed that the highest transfection efficiency would be obtained when cell fusion degree was 81%-90%,liposome volume was 1.8 μl,liposome/plasmid ratio was 2∶1 and transfecting time was 4 h.[Conclusion] The study will provide a test basis for the construction of mammary gland bioreactors.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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