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
Effect of vip on the glycogen metabolism of human colon adenocarcinoma cells studied by 13C nuclear magnetic resonance spectroscopy
โ Scribed by Jean-Philippe Galons; Jacques Fantini; Jean Vion-Dury; Patrick J. Cozzone; Paul Canioni
- Publisher
- John Wiley and Sons
- Year
- 1990
- Tongue
- French
- Weight
- 661 KB
- Volume
- 45
- Category
- Article
- ISSN
- 0020-7136
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โฆ Synopsis
Metabolic pathways of glucose utilization have been investigated in a human colon adenocarcinoma cell line (HT29) using carbon-I3 Nuclear Magnetic Resonance spectroscopy. HT29 cells were adapted to grow on a polystyrene beaded microcarrier and were perfused when attached to the beads in a specially designed NMR cell. Abnormalities in carbohydrate metabolism already observed in several cancer cells were studied in HT29 cells fed with (I -%)-enriched glucose. The cells were first perfused with a glucose-free medium for 2 hr in order to deplete the intracellular store of glycogen, and they were subsequently perfused with a medium containing enriched glucose at an initial concentration of 5.5 mM. Sequential I3CC-NMR spectra, recorded at 100.5 MHz (5 min accumulation), show that HT29 cells were able to utilize glucose through the glycolytic pathway while storing glucose as glycogen (glucose was utilized at a rate of 3.9 pmol/mg protein/ hr). The glycolytic activity determined by the amount of lactic acid produced was 4.6 pdmg proteidhr, corresponding to the formation of 1.2 lactic acid per glucose molecule. Glycogen accumulation corresponded to 16 pg/mg of protein. Treatment of HT29 with 10 nM vasoactive intestinal peptide (VIP) induced a transient decrease in the level of labelled glycogen to 50% of the initial value. Control level was recovered I 2 min after VIP loading.
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