Tissue Culture Media 

 Reagents, Difco 



As IN the field of dehydrated culture media for Bacteriology and Mycology, 

 Difco Laboratories has pioneered in the preparation of dehydrated reagents 

 for in vitro propagation and maintenance of tissue cells. The task of preparing 

 and distributing these highly specialized products was undertaken at the request 

 of the Tissue Culture Association. Each of these reagents, before being released 

 for distribution, is tested and certified for use in tissue culture by the Central 

 Laboratory of the Tissue Culture Association. They are applicable to the slide, 

 roller tube and flask culture techniques commonly employed for propagation and 

 study of tissue cells in vitro. All desiccated reagents are dried from the frozen 

 state to preserve, unaltered, the properties of the original material. The reagents 

 currently available are those commonly employed for culturing tissue cells, 

 maintaining tissue banks, and for the propagation of Viruses. Additional reagents 

 will be made available as required for these and other procedures related to 

 tissue culture. 



TG CHICKEN PLASMA (B354) 



TG Chicken Plasma is sterile desiccated whole chicken plasma certified for 

 use in culturing tissue cells in vitro. It is prepared from selected disease-free 

 cockerels 4-8 months old and is packaged in 5 ml. quantities. 



TC Chicken Plasma is reconstituted to its original volume by adding, asep- 

 tically, 4.5 ml. sterile TC Reconstituting Fluid or TC Triple Distilled Water to 

 each bottle. The Reconstituting Fluid may be added conveniently with a sterile 

 5 ml. syringe. Solution of the material is accelerated by gentle end-over-end rota- 

 tion of the bottle. Do not shake. Allow 30 minutes for complete solution of the 

 desiccated plasma. 



The reconstituted plasma is alkaline in reaction due to loss of carbon dioxide 

 in the desiccation process and requires adjustment with carbon dioxide to pH 

 7.2-7.4 if a normal clotting time is desired. Adjustment of the reaction with 

 carbon dioxide to pH 7.2-7.4 accelerates the clotting time when Embryo Extract 

 and tissue explants are added. The adjustment of the reaction of the recon- 

 stituted plasma is best accomplished by first adding TC Phenol Red Solution 

 1% to the plasma to give a final Phenol Red concentration of 0.002 per cent, 

 (add 1 ml. TC Phenol Red Solution 1% to 9 ml. TC Triple Distilled Water, then 

 add 0.1 ml. of resultant Phenol Red Solution to 5 ml. of reconstituted Chicken 

 Plasma) and then introducing carbon dioxide to bring the reaction to pH 

 7.2-7.4. Three methods of introducing carbon dioxide that have given good 

 results are outlined: 



1. Transfer the reconstituted TC Chicken Plasma containing 0.002 per cent 

 Phenol Red to a clean, dry, sterile cotton stoppered or loosely capped test tube. 

 Place the tube in a beaker or container of sufficient depth so that the walls ex- 

 tend one or more inches above the mouth of the tube. Introduce carbon dioxide 

 into the beaker to displace the air and then cover the beaker with paper or 

 suitable cover. Gently agitate the plasma in the tube occasionally to bring all 

 parts of the solution in contact with the carbon dioxide. Observe color change in 



301 



