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An efficient method for routine epstein-barr virus immortalization of human B lymphocytes |
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| Authors: | Florence E. Wall Richard D. Henkel Michael P. Stern Hal B. Jenson Mary Pat Moyer |
| Affiliation: | (1) Center for Human Cell Biotechnology, Department of Surgery, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, 78284-7842 San Antonio, Texas;(2) Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, Texas;(3) Department of Medicine, The University of Texas Health Science Center at San Antonio, 78284, Texas;(4) Department of Pediatrics, The University of Texas Health Science Center at San Antonio, 78284, Texas |
| Abstract: | Summary A variety of methods exist for the immortalization of B lymphocytes by Epstein-Barr virus due to the simplicity of such techniques to establish cell lines with stable genomic DNA. Two different methods for immortalizing lymphoblastoid cell lines were compared for differences in techniques and materials, time between initiation and immortalization, and success rate of immortalization. An incubation period in Epstein-Barr virus and the use of conditioned media improved immortalization efficiency from 86 to 98% and decreased the time (usually weeks) from culture initiation to cryopreservation. The resulting cell bank was used to produce DNA for genetic studies focusing on the genes involved in non-insulin-dependent diabetes mellitus. |
| Keywords: | immortalization B lymphocytes Epstein-Barr virus transformation human cell lines |
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