Cell Cycle Analysis as a Tool for Control and Regulation of Mammalian Cell Cultures in Bioreactors

Deutschmann, Sven; Valley, Ulrich; Jäger, Volker; Wagner, Roland

doi:10.1016/b978-0-7506-1845-8.50087-9

Cited by 1 publication

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“…Nucleotide ratios Ryll and Wagner (1992) proposed the so-called NTP-U plot as a useful tool for process control which has been later successfully applied in different mammalian cell culture processes (Valley et al 1993;Deutschmann et al 1994;Grammatikos et al 1999). Here, we first used the NTP-U plot to characterize dedifferentiation of in vitro-cultured primary porcine hepatocytes.…”

Section: Albumin Secretionmentioning

confidence: 99%

Intracellular nucleotide pools and ratios as tools for monitoring dedifferentiation of primary porcine hepatocytes in culture

Rocker

Hesse²,

Bader

et al. 2006

Cytotechnology

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The effect of two culture configurations (single collagen gel and double collagen gel) and of two hormones (insulin and glucagon) on the differentiated status and the intracellular nucleotide pools of primary porcine hepatocytes was investigated. The objective was to analyze and monitor the current state of differentiation supported by the two culture modes using intracellular nucleotide analysis. Specific intracellular nucleotide ratios, namely the nucleoside triphosphate (NTP) and the uridine (U) ratio were shown to consistently reflect the state of dedifferentiation status of the primary cells in culture affected by the presence of the two hormones insulin and glucagon. Continuous dedifferentiation of the cells was monitored in parallel by the reduction of the secretion of albumin, and changes in UDP-activated hexoses and UDPglucuronate. The presence of insulin maintained the differentiated status of hepatocytes for more than 12 days when cultivated under double gel conditions whereas glucagon was less effective. In contrast, cells cultivated in a single gel matrix immediately started to dedifferentiate upon seeding. NTP and U ratios were shown to be more sensitive for monitoring dedifferentiation in culture than the albumin secretion. Their use allowed the generation of an easily applicable NTP-U plot in order to give a direct graphical representation of the current differentiation status of the cultured cells. Moreover, the transition from functional and differentiated hepatocytes to dedifferentiated fibroblasts could be determined earlier by the nucleotide ratios compared to the conventional method of monitoring the albumin secretion rate.

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