Sonderhoff S A, Kilburn D G, Piret J M
Department of Microbiology, University of British Columbia, Vancouver, British Columbia, Canada.
Biotechnol Bioeng. 1992 Apr 5;39(8):859-64. doi: 10.1002/bit.260390807.
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.
在杂交瘤细胞培养中，研究了将细胞ATP分析作为测量活生物质负载量的一种方法。通过荧光素-荧光素酶测定法进行的ATP分析与台盼蓝染色血细胞计数器计数进行了比较。在分批培养过程中，每个活细胞的细胞特异性ATP含量在2至6飞摩尔之间变化。ATP水平在指数生长期最高，在稳定期和衰退期下降。进行电子计数和体积测量以测定活细胞生物量。使用荧光素二乙酸酯进行细胞分选，以分离活细胞率在35%至90%之间的培养物中的活细胞和非活细胞。活细胞的细胞特异性ATP比非活细胞高两个数量级以上。细胞特异性ATP与活细胞体积直接相关，而不是与活细胞数量相关。在分批培养条件范围内，ATP分析应比分批血细胞计数器计数更准确地测量杂交瘤活生物质。
