Growth of a human colonic adenocarcinoma cell line (HT 29) on microcarrier beads: Metabolic studies by 31phosphorus nuclear magnetic resonance spectroscopy

Jacques Fantini Baetselier, Jacques Marvaldi, Jean‐Philippe ‐P Galons, Patrick J. Cozzone, Paul Canioni

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

A method allowing the growth of a human colon adenocarcinoma cell line (HT 29) on beaded polystyrene microcarriers has been developed by modifying the culture conditions used in monolayer cultures. Under optimized conditions, the cells became confluent 7 days after seeding and reached a density of 2.8 x 105 cells/cm of microcarrier (65% of the available area occupied). 31P NMR spectra were typically recorded on 300 x 106 cells continuously perfused at a flow rate of 15 ml/min in a specially designed NMR chamber in which the micro‐carrier beads were sequestered within the receiver coil volume. The in vivo spectrum displays a series of resonances assigned to nucleoside triphosphates (ATP and GTP), inorganic phosphate and various phosphomonoesters (mainly glucose‐6‐P and phosphorylcholine). Diphosphodiester resonances (DPDE, mainly UDP‐N‐acetyl‐glucosamine and UDP‐N‐acetylgalactosamine) were not detected in the in vivo spectrum and were only apparent in the spectrum of the perchloric acid extract of the cells, indicating that these compounds have a restricted mobility in the intracellular compartment. The intracellular pH of HT 29 cells was 7.2 during the perfusion with a medium buffered at pH 7.3. The internal pH decreased slowly (2 x 10‐3 pH unit/min) during anoxic perfusion, but severe intracellular acidosis occurred after 40 min of ischemia (2.7 x 10‐1 pH unit/min). Sequential recording of 31P NMR spectra has shown that HT 29 cells are able to maintain their high energy phosphorylated compound levels (ATP) when subjected to 100 min of anoxia and 40 min of total ischemia.

Original languageEnglish (US)
Pages (from-to)255-260
Number of pages6
JournalInternational Journal of Cancer
Volume39
Issue number2
DOIs
StatePublished - Feb 15 1987
Externally publishedYes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Fingerprint Dive into the research topics of 'Growth of a human colonic adenocarcinoma cell line (HT 29) on microcarrier beads: Metabolic studies by <sup>31</sup>phosphorus nuclear magnetic resonance spectroscopy'. Together they form a unique fingerprint.

  • Cite this