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Analysis of mammalian viable cell biomass based on cellular ATP
Author(s) -
Sonderhoff Stefan A.,
Kilburn Douglas G.,
Piret James M.
Publication year - 1992
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260390807
Subject(s) - hemocytometer , trypan blue , cell culture , cell counting , biology , cell , cell growth , fluorescein , cell sorting , biochemistry , microbiology and biotechnology , chemistry , cell cycle , genetics , physics , quantum mechanics , fluorescence
Analysis of cellular ATP as a means of measuring viable biomass loading was investigated in hybridoma cell culture. ATP analysis by the luciferin–luciferase assay was compared with trypan blue‐stained hemocytometer counts. The cell‐specific ATP content varied between 2 and 6 fmol per viable cell over a batch culture. ATP levels were highest during exponential growth, and decreased during the stationary and decline phases. Electronic counting and volume measurements were performed to assay the viable cell biomass. Cell sorting, using fluorescein diacetate, was used to separate viable and nonviable cells in cultures with between 35% and 90% viable cells. Viable cells contained over 2 orders of magnitude greater cell‐specific ATP than nonviable cells. Cell‐specific ATP correlated directly with the viable cell volume rather than viable cell numbers. Over the range of batch culture conditions, ATP analysis should provide a more accurate measurement of hybridoma viable biomass than hemocytometer counts.