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Construction of NIH3T3-CD40L Stable Cell Lines and Its Application in EBV Transformation of B Lymphocytes
Liu Xi, Zhao Ye, Xu Chongfeng, Duan Ziyuan*
Genetic Resource Research Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
Abstract: Stably expressing CD40L NIH3T3 cell lines were constructed and its application as feeder cells to culture and establish B lymphoblastoid cell lines (B-LCLs) in vitro were evaluated. pCMV6-CD40L was transfected into NIH3T3 cell lines through lipofectamine method， followed NIH3T3 that express CD40L were screened by G418. Positive clones were confirmed at RNA and protein level， the NIH3T3-CD40L cell lines were used as feeder cells in culturing B-LCLs and in the process of EBV transformation. The results showed that NIH3T3-CD40L cell lines increased the survival rate and promote the proliferation of B-LCLs in low density and reduce the cell concentration required to establish B-LCLs. Therefore， NIH3T3-CD40L cell lines have been successfully constructed which would lay a foundation for further studies， such as monoclonal antibody production.
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