Adenosine phosphorylase activity in mycoplasma-free growth media for mammalian cells.
Mammalian cells have enzymes that deaminate adenosine to inosine, which can readily be phosphorolysed to hypoxanthine. They do not, however, possess enzymes to form adenine by the cleavage of adenosine. For this reason, the release of adenine from adenosine by mammalian cell cultures has usually been interpreted as indicating the presence of mycoplasma, a frequent microbial contaminant that contains high levels of adenosine phosphorylase. We found that some human lymphoblast cultures free of mycoplasma showed high levels of adenosine cleavage and that this activity resulted from adenosine phosphorylase in the bovine serum used as the culture growth supplement. A survey of 13 serum supplements disclosed that fetal bovine serum (six lots) contains the highest adenosine phosphorylase activity, ranging from 9 to 648 nmol adenine produced per hour per ml serum; newborn calf serum (four lots) has much less activity, ranging from 0 to 5 nmol adenine produced per hour per ml serum; and donor horse serum (three lots) contains no detectable activity. These results suggest that mycoplasma tests dependent on the presence of adenosine phosphorylase or other enzyme activities may give false-positives with cultures containing fetal bovine serum supplements.
D000241:Adenosine / Q000378:metabolism; D000818:Animals; D001769:Blood; D002417:Cattle / Q000097:blood; D002460:Cell Line; D003470:Culture Media / Q000032:analysis*; D003839:Deoxyadenosines / Q000378:metabolism; D006801:Humans; D008214:Lymphocytes; D009174:Mycoplasma / Q000254:growth & development; D010430:Pentosyltransferases / Q000097:blood*; D011683:Purine-Nucleoside Phosphorylase / Q000097:blood*
D003470:Culture Media; D003839:Deoxyadenosines; D010430:Pentosyltransferases; C019603:adenosine phosphorylase; D011683:Purine-Nucleoside Phosphorylase; D000241:Adenosine