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LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

Appendix

REFERENCE NO: AHC/1998/3/1.2 Appendix 7


TITLE: PREPARATION OF GROWTH MEDIUM FACTOR SUPPLEMENT (GMF)


INTRODUCTION

This growth supplement should be added to poorly growing lymphoblastoid lines at any stage of transformation. See COSHH form LEB06.

PROCEDURE

  1. Mix all the ingredients below together in a 15ml sterile tube:
  • 2-mercaptoethanol (50mM): 0.650ml
  • non-essential amino acids, supplied 100 x concentrate: 5.5ml
  • Sodium selenite (2 x 10-6M): 0.250ml
  • Transferrin (4mg/ml): 0.250ml
  • Sodium pyruvate supplied 100 x concentrate: 5.5ml

TOTAL 12.15ml

  1. Sterile filter with a 2mm filter and dispense aliquots into labelled cryotubes.
  2. Record the preparation in the Media Preparation Book and assign the next number, tubes should be labelled accordingly.
  3. Mark this number on each vial as well as GMF, and store at <-10C in freezer.
  4. Thaw as for thawing cells, and add to 500 ml of medium when required.


Guidelines prepared for CABRI by CERDIC, DSMZ, ECACC, INRC, November 1998
Page layout by CERDIC
Copyright CABRI, 1998

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