Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



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Preparing Fibroblast Feeder Cell Cultures

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Certain fibroblast cultures secrete the necessary factors to allow the growth of hybridoma cells at low plating densities. Early studies used fibroblast cultures that had been treated with mitomycin C or lethal doses of irradiation. Both of these treatments made it impossible for the feeder cells to contaminate future cultures of the hybridomas. More recently, this has been shown not to be necessary for fibroblast cultures that adhere strongly to the plastic tissue culture surface. Other studies have compared the ability of different fibroblast cells to support single-cell cultures of hybridoma cells and have found that the human diploid cells MRC-5 are the most effective in this test. These cells are not an established cell line and thus will need to be replaced in the future by another source. The MRC-5 cells (live or pre-irradiated) are currently available from several sources including the American Type Culture Collection (ATCC).

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

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