Development of a serum-free co-culture of human intestinal epithelium cell-lines (Caco-2/HT29-5M21)

in BMC Cell Biology (2006), 7

Background: The absorptive and goblet cells are the main cellular types encountered in the intestine epithelium. The cell lineage Caco-2 is a model commonly used to reproduce the features of the bowel ... [more ▼]

Background: The absorptive and goblet cells are the main cellular types encountered in the intestine epithelium. The cell lineage Caco-2 is a model commonly used to reproduce the features of the bowel epithelium. However, there is a strong debate regarding the value of Caco-2 cell culture to mimick in vivo situation. Indeed, some authors report in Caco-2 a low paracellular permeability and an ease of access of highly diffusible small molecules to the microvilli, due to an almost complete lack of mucus. The HT29-5M21 intestinal cell lineage is a mucin-secreting cellular population. A co-culture system carried out in a serum-free medium and comprising both Caco-2 and HT29-5M21 cells was developed. The systematic use of a co-culture system requires the characterization of the monolayer under a given experimental procedure. Results: In this study, we investigated the activity and localization of the alkaline phosphatase and the expression of IAP and MUC5AC genes to determine a correlation between these markers and the cellular composition of a differentiated monolayer obtained from a mixture of Caco-2 and HT29-5M21 cells. We observed that the culture conditions used ( serum-free medium) did not change the phenotype of each cell type, and produced a reproducible model. The alkaline phosphatase expression characterizing Caco-2 cells was influenced by the presence of HT29-5M21 cells. Conclusion: The culture formed by 75% Caco-2 and 25% HT29-5M21 produce a monolayer containing the two main cell types of human intestinal epithelium and characterized by a reduced permeability to macromolecules. [less ▲]

Postnatal maturation of intestine and spleen, induced by polyamines in suckling rats

in Morgan, D.; Bauer, F.; White, A. (Eds.) COST 917 Biologically active amines in food. Volume VII (2005)

Polyamines in gut inflammation and allergy

in Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry (2004), 3

The natural polyamines, named 1,4-diaminobutane, N-aminopropyl-1,4-diaminobutane and N,N'-bisaminopropyl- 1,4-diaminobutane, are also designated respectively as putrescine, spermidine and spermine. They ... [more ▼]

The natural polyamines, named 1,4-diaminobutane, N-aminopropyl-1,4-diaminobutane and N,N'-bisaminopropyl- 1,4-diaminobutane, are also designated respectively as putrescine, spermidine and spermine. They are polycationic compounds found in all eukaryotic cells. As they are deeply involved in cell functions, e.g. cellular growth, their concentration and their equilibrium in the intracellular pool are maintained in very narrow limits by regulatory mechanisms acting in a very fast, sensitive and precise way. These compounds are involved in gut inflammatory processes and in allergy. Indeed, they control the production of inflammatory mediators in several cell lines or tissues. Polyamine metabolism could be considered as a putative target for inflammation, allergy prevention or therapy. [less ▲]

Polyamines as essential substances in metabolism

in Trends in Comparative Biochemistry and Physiology (2002), 9

Are Milk Polyamines Preventive Agents against Food Allergy?

in Proceedings of the Nutrition Society (2000), 59(1), 81-6

Insufficient polyamine intake could play a role in the induction of sensitization to dietary allergens. This proposal is based essentially on investigations made in sucking rats and in children. In ... [more ▼]

Insufficient polyamine intake could play a role in the induction of sensitization to dietary allergens. This proposal is based essentially on investigations made in sucking rats and in children. In sucking rats it has been established that oral administration of spermine can induce all the modifications occurring in the digestive tract at weaning. In the intestine events occur in two phases. The early event consists of desquamation of the epithelium resulting from an activation of apoptosis. The late event appears to involve an hormonal cascade in which adrenocorticotropic hormone, cytokines, bombesin and corticosterone are included. Observations in human subjects show that: (1) the spermine and spermidine concentrations are generally lower in infant formulas than in human breast milk. Mothers seem consistently to have relatively high or relatively low concentrations of spermine and spermidine in their milk. These individual variations may be due to diet, lifestyle or genetic background; (2) the probability of developing allergy can reach 80 % if the mean spermine concentration in the milk is lower than 2 nmol/ml milk. It is approximately 0 % if the mean spermine concentration is higher than 13 nmol/ml milk; (3) preliminary results show that the intestinal permeability to macromolecules differs in premature babies when they are fed on breast milk compared with infant formulas (J Senterre, J Rigo, G Forget, G Dandrifosse and N Romain, unpublished results). This difference does not seem to be present when powdered milk is supplemented with polyamines at the concentration found in breast milk; (4) spermine increases proliferation and differentiation of lymphocytes isolated from the tonsils of children. [less ▲]

Polycations : relationship between chemical structure and biological behavior

in Morgan, D.; White, A.; Sanchez-Jimenez, F. (Eds.) et al COST 917 Biologically active amines in food. Volume IV (2000)

Effects of a long-term low-polyamine diet on biochemical parameters and polyamine content in different organs of the germ-free rat

in Bardocz, S.; White, A.; Hajos, G. (Eds.) COST 917 Biogenically active amines in food. Volume II : Metabolic effects of biologically active amines in food (1998)

Role of Interleukin-1 Beta, Interleukin-6, and Tnf-Alpha in Intestinal Maturation Induced by Dietary Spermine in Rats

in Endocrine (1997), 6(2), 187-94

In the present investigation, the authors aimed to evaluate the role of cytokines in intestinal postnatal maturation induced by dietary polyamines. Neonatal rats were administered either saline (8 mumol ... [more ▼]

In the present investigation, the authors aimed to evaluate the role of cytokines in intestinal postnatal maturation induced by dietary polyamines. Neonatal rats were administered either saline (8 mumol) orally. Spermine increased interleukin-1 beta (IL-1 beta), IL-6, and TNF-alpha plasma concentration. The maximum concentrations of IL-1 beta, IL-6, and TNF-alpha were, respectively, observed at 4, 4, and 8 h posttreatment. Intraperitoneal (i.p.) injection of IL-1 beta increased the specific activity of sucrase in whole small intestine, whereas the specific activities of maltase and lactase were significantly enhanced only in the jejunum. IL-6 elicited sucrase and increased maltase specific activity in the whole small intestine, but lactase specific activity was not affected. TNF-alpha had no effect on sucrase and maltase specific activity, but a slight augmentation of lactase specific activity was detected in the jejunum. Spermine and spermidine content in the intestine was increased by i.p. injection of IL-1 beta and IL-6. Corticosterone secretion was elevated by single i.p. injection of IL-1 beta, IL-6, or TNF-alpha. These findings suggest that spermine could induce postnatal intestinal development and corticosterone secretion through a cytokine-dependent mechanism. [less ▲]