Cytotechnology

, Volume 17, Issue 3, pp 193–202 | Cite as

The culture of rat myeloma and rat hybridoma cells in a protein-free medium

  • M. J. Keen
Technical Report

Abstract

Y0 is a rat x rat hybridoma cell line, which does not secrete immunoglobulin, produced using a fusion partner derived from the Y3 (Y3,Ag.1.2.3) rat myoloma cell line. Y0 and Y3 have both been widely used as fusion partners in the production of rat x rat hybridomas. Y0 has also been used in recombinant gene technology. Y0 cells grown in shake flask culture, using RPMI 1640 medium with 4mM l-glutamine and 5% foetal bovine serum, reached a maximal cell density of 1.5×106 cells ml−1 with 86% viability. Y0 cells which has been adapted to grow in ABC protein-free medium reached a maximal density, in shake flask culture, of 8.75×105 cells ml−1 with 79% viability. An improved protein-free medium, designated W38 medium, was developed. In shake flask culture, W38 medium supported Y0 cell growth to a density of 2.02×106 cells ml−1 with 96% viability. Two Y3 hybridomas, YID 13.9.4 cells and SAM 618 cells were adapted to growth in W38 medium. For both hybridomas, cell growth and product yield in shake flask culture using W38 medium was superior to that obtained with serum-containing RPMI 1640 medium.

Key words

Y0 hybridoma myeloma protein-free culture medium 

Abbreviations

F12

Hams F12 medium

DMEM

Dulbeccos medium

RPMI

RPMI 1640 medium

FBS

foetal bovine serum

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • M. J. Keen
    • 1
  1. 1.Biology Research DivisionWellcome Research LaboratoiresBeckenhamUK

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