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LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

Appendix

REFERENCE NO: AHC/1998/3/1.2 Appendix 6


TITLE: PREPARATION OF HB SUPPLEMENT


INTRODUCTION

This media supplement may be added to tissue culture media for slow growing cell lines such as hybridomas.

 

PROCEDURE

Materials

  • 1320 mg oxaloacetic acid
  • 80 mg bovine insulin
  • 550 mg sodium pyruvate
  • 100 labelled and dated 1 ml cryovials
  • 0.2 mm filters, 20 ml syringes
  1. Add the oxaloacetic acid and bovine insulin to 80 ml of distilled water.
  2. Dissolve in a 37C water bath at between 35 to 38C.
  3. Add 550 mg of sodium pyruvate.
  4. Adjust to 100 ml final volume with dH2O.
  5. Filter sterilise, aliquot and store at <-10C.

The solution is used at 1:100 final dilution in media. Mix well on thawing as precipitates may form and add to tissue culture medium.

 


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

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