Over the last 5 years, the ATP cell viability assay (ATP-CVA) has been
used to study the in vitro response of cell lines and fresh gynecolog
ic human tumors to a variety of antineoplastic agents including chemot
herapeutic agents, hormones and biological response modifiers. This as
say measures light production as intracellular ATP interacts with the
luciferin-luciferase complex. Quantitation of the light produced has b
een shown to directly correspond with the number of viable cells. A pa
st criticism is that in the ATP-CVA, when applied to fresh tumor tissu
e, normal cells (fibroblasts, macrophages and lymphocytes) also produc
e ATP, and if present in sufficient numbers, could lead to errors in c
hemosensitivity testing results. This study was designed to evaluate t
he growth characteristics of various benign cells found in fresh tumor
s. The cells were studied under multiple plating conditions to show th
e relative increase or decrease of fractional ATP measured at differen
t time points. We found that agar/McCoy underlayer and agarose-coated
wells do not permit the growth of nonmalignant cells. In the culture c
onditions of the ATP-CVA, nonmalignant cells do not contribute relevan
t ATP levels when treated samples are compared to controls on day 6. T
herefore, results of the ATP-CVA in fresh tumors should not be affecte
d.