An approach to a selection system for adenosine-deaminase-positive (ADA+) cells and detection of rat ADA+ "revertants".
We have substituted deoxyadenosine or adenosine for hypoxanthine in the standard HAT selection system in an attempt to select for ADA-normal (ADA+) cells. ADA- human lymphoid line cells could not utilize deoxyadenosine as an alternative to hypoxanthine as a purine source (DAT) and failed to grow but were only somewhat inhibited in growth when adenosine was substituted for hypoxanthine (AAT). In contrast, ADA+ cells utilized adenosine or deoxyadenosine as efficiently as hypoxanthine as a purine source. Growth in DAT, but not in HAT, of an artificial mixture of one ADA+ human lymphoid cells in 1,000 ADA- cells resulted in enrichment of ADA+ cells to 25-86% of total cells. When we grew a rat ADA- cell line in two variations of the DAT system, we detected at least three electrophoretically different ADA+ patterns, one of which corresponded to normal rat ADA. These could represent "revertants."
D000241:Adenosine / Q000378:metabolism* / Q000494:pharmacology; D000243:Adenosine Deaminase / Q000235:genetics* / Q000378:metabolism; D000630:Aminopterin / Q000494:pharmacology; D000818:Animals; D002455:Cell Division; D002459:Cell Fusion; D002460:Cell Line; D002470:Cell Survival; D003470:Culture Media; D003839:Deoxyadenosines / Q000378:metabolism* / Q000494:pharmacology; D004592:Electrophoresis, Starch Gel; D006801:Humans; D009154:Mutation; D009700:Nucleoside Deaminases / Q000235:genetics*; D051381:Rats; D012641:Selection, Genetic; D013936:Thymidine / Q000494:pharmacology
D003470:Culture Media; D003839:Deoxyadenosines; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase; D000630:Aminopterin; D000241:Adenosine; D013936:Thymidine