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Improving the long‐term storage of a mammalian biosensor cell line via genetic engineering
Authors:Martha S. Petrovick  Frances E. Nargi  Theresa Towle  Kristine Hogan  Marie Bohane  Deborah J. Wright  Thomas H. MacRae  Malcolm Potts  Richard F. Helm
Affiliation:1. MIT Lincoln Laboratory, Lexington, Massachusetts 024 20;2. telephone: 781‐981‐1995;3. fax: 781‐981‐3867;4. Department of Biochemistry, Virginia Tech, Blacksburg, Virginia;5. Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada
Abstract:The unique properties of mammalian cells make them valuable for a variety of applications in medicine, industry, and diagnostics. However, the utility of such cells is restricted due to the difficulty in storing them non‐frozen for an extended time and still maintaining their stability and responsiveness. In order to extend the active life span of a mammalian biosensor cell line at room and refrigerated temperatures, we have over expressed genes that are reported to provide protection from apoptosis, stress, or oxidation. We demonstrated that over expression of genes from the extremophile, Artemia franciscana, as well as GADD45β, extends room‐temperature storage of fully active cells 3.5‐fold, while over production of several anti‐apoptotic proteins extended 4°C storage 2‐ to 3‐fold. Methodologies like these that improve the stability of mammalian‐cell‐based technologies in the absence of freezers may enable widespread use of these tools in applications that have been considered impractical based solely on limited storage characteristics. Biotechnol. Bioeng. 2010; 106: 474–481. © 2010 Wiley Periodicals, Inc.
Keywords:apoptosis  anti‐apoptosis  mammalian cells  cell engineering  extremophiles  cell‐based assays
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