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
- 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
- Sterile filter with a 2mm filter and dispense aliquots into labelled cryotubes.
- Record the preparation in the Media Preparation Book and assign the next number, tubes should be labelled accordingly.
- Mark this number on each vial as well as GMF, and store at <-10°C in freezer.
- 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
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