Advantages, Disadvantages and Limitations of the System

TNO has 10 units of TIM-2 available, allowing multiple parameters to be tested in parallel. In contrast to other multi-compartmental in vitro models, experiments are quick (usually three test days or less), yet still physiological and predictive. The advantages over other models are the presence of peristaltics and a dialysis system, the latter of which allows production of physiological concentrations of metabolites, and a highly active microbiota of normal density to be used. Another advantage is that a single parameter in the system can be changed, and the effect of that single parameter on microbiota activity can be studied. This has been done e.g., when studying the effect of different pH's on fermentation of carbohydrates (unpublished). Naturally, in vitro models have their limitations. As with every other in vitro model that mimics the colon, TIM-2 does not have epithelial or immune cells. However, as discussed in Sect. 26.4, samples can be incubated with these cells for even better predictability. Another limitation is that the model has been developed on the basis of literature data of mostly health individuals. Due to this, it is unclear exactly which parameters to simulate when simulating patient populations as discussed in van Nuenen et al. (2004). Another limitation is that (apart from volume and pH) there are no feed-back mechanisms in the system. Therefore, the experiments in such in vitro models will always be at most an indication of what may occur in real life, and the results need to be interpreted with care.


Aguirre A, Ramiro-Garcia J, Koenen ME, Venema K (2014) To pool or not to pool? Impact of the use of individual and pooled faecal samples for in vitro fermentation studies. J Microbiol Methods 107:1–7

Binsl TW, De Graaf AA, Venema K, Heringa J, Maathuis A, De Waard P, Van Beek JH (2010) Measuring non-steady-state metabolic fluxes in starch-converting faecal microbiota in vitro. Benefic Microbes 1:391–405

Bordonaro M, Venema K, Putri AK, Lazarova D (2014) Approaches that ascertain the role of dietary compounds in colonic cancer cells. World J Gastrointest Oncol 6:1–10

Bussolo CS, Roeselers G, Troost F, Jonkers D, Koenen ME, Venema K (2014) Prebiotic effects of cassava bagasse in TNO's in vitro model of the colon (TIM-2) in lean versus obese microbiota.

J Funct Foods 11:210–220

den Besten G, van Eunen K, Groen AK, Venema K, Reijngoud DJ, Bakker BM (2013) The role of short-chain fatty acids in the interplay between diet, gut microbiota, and host energy metabolism. J Lipid Res 54:2325–2340

Gao K, Xu A, Krul C, Venema K, Liu Y, Niu Y, Lu J, Bensoussan L, Seeram NP, Heber D, Henning SM (2006) Of the major phenolic acids formed during human microbial fermentation of tea, citrus, and soy flavonoid supplements, only 3,4-dihydroxyphenylacetic acid has antiproliferative activity. J Nutr 136:52–57

Havenaar R (2011) Intestinal health functions of colonic microbial metabolites: a review. Benefic Microbes 2:103–114

Kong H, Wang M, Venema K, Maathuis A, van der Heijden R, van der Greef J, Xu G, Hankemeier T (2009) Bioconversion of red ginseng saponins in the gastro-intestinal tract in vitro model studied by high-performance liquid chromatography-high resolution Fourier transform ion cyclotron resonance mass spectrometry. J Chromatogr A 1216:2195–2203

Kovatcheva-Datchary P, Egert M, Maathuis A, Rajilic-Stojanovic M, de Graaf AA, Smidt H, de Vos WM, Venema K (2009) Linking phylogenetic identities of bacteria to starch fermentation in an in vitro model of the large intestine by RNA-based stable isotope probing. Environ Microbiol 11:914–926

Lamers RJ, Wessels EC, van de Sandt JJ, Venema K, Schaafsma G, van der Greef J, van Nesselrooij JH (2003) A pilot study to investigate effects of inulin on Caco-2 cells through in vitro metabolic fingerprinting. J Nutr 133:3080–3084

Maathuis AJ, van den Heuvel EG, Schoterman MH, Venema K (2012) Galacto-oligosaccharides have prebiotic activity in a dynamic in vitro colon model using a (13)C-labeling technique. J Nutr 142:1205–1212

Macfarlane GT, Gibson GR, Cummings JH (1992) Comparison of fermentation reactions in different regions of the human colon. J Appl Bacteriol 72:57–64

Martinez RC, Cardarelli HR, Borst W, Albrecht S, Schols H, Gutierrez OP, Maathuis AJ, de Melo Franco BD, De Martinis EC, Zoetendal EG, Venema K, Saad SM, Smidt H (2012) Effect of galactooligosaccharides and Bifidobacterium animalis Bb-12 on growth of Lactobacillus amylovorus DSM 16698, microbial community structure, and metabolite production in an in vitro colonic model set up with human or pig microbiota. FEMS Microbiol Ecol 84:110–123

Minekus M, Smeets-Peeters M, Bernalier A, Marol-Bonnin S, Havenaar R, Marteau P, Alric M, Fonty G, Huis in't Veld JH (1999) A computer-controlled system to simulate conditions of the large intestine with peristaltic mixing, water absorption and absorption of fermentation products. Appl Microbiol Biotechnol 53:108–114

Rajilic-Stojanovic M, Maathuis A, Heilig HG, Venema K, de Vos WM, Smidt H (2010) Evaluating the microbial diversity of an in vitro model of the human large intestine by phylogenetic microarray analysis. Microbiology 156:3270–3281

Ramasamy US, Venema K, Schols HA, Gruppen H (2014) The effect of soluble and insoluble fibers within the in vitro fermentation of chicory root pulp by human gut bacteria. J Agric Food Chem 62(28):6794–6802

Rehman A, Heinsen FA, Koenen ME, Venema K, Knecht H, Hellmig S, Schreiber S, Ott SJ (2012) Effects of probiotics and antibiotics on the intestinal homeostasis in a computer controlled model of the large intestine. BMC Microbiol 12:47

Robert C, Bernalier-Donadille A (2003) The cellulolytic microflora of the human colon: evidence of microcrystalline cellulose-degrading bacteria in methane-excreting subjects. FEMS Microbiol Ecol 46:81–89

Roeselers G, Bouwman J, Venema K, Montijn R (2012) The human gastrointestinal microbiota-an unexplored frontier for pharmaceutical discovery. Pharmacol Res 66:443–447

Rose DJ, Venema K, Keshavarzian A, Hamaker BR (2010) Starch-entrapped microspheres show a beneficial fermentation profile and decrease in potentially harmful bacteria during in vitro fermentation in faecal microbiota obtained from patients with inflammatory bowel disease. Br J Nutr 103:1514–1524

Slavin JL, Brauer PM, Marlett JA (1981) Neutral detergent fiber, hemicellulose and cellulose digestibility in human subjects. J Nutr 111:287–297

van Nuenen MHMC, Meyer PD, Venema K (2003) The effect of various inulins and Clostridium difficile on the metabolic activity of the human colonic microbiota in vitro. Microb Ecol Health Dis 15:137–144

van Nuenen MH, Venema K, van der Woude JC, Kuipers EJ (2004) The metabolic activity of fecal microbiota from healthy individuals and patients with inflammatory bowel disease. Dig Dis Sci 49:485–491

van Nuenen MH, de Ligt RA, Doornbos RP, van der Woude JC, Kuipers EJ, Venema K (2005) The influence of microbial metabolites on human intestinal epithelial cells and macrophages in vitro. FEMS Immunol Med Microbiol 45:183–189

Venema K (2010) Role of gut microbiota in the control of energy and carbohydrate metabolism.

Curr Opin Clin Nutr Metab Care 13:432–438

Venema K (2011) Stable isotope probing and the human gut. In: Murrell JC, Whiteley AS (eds) Stable isotope probing and related technologies. ASM Press, Washington, DC, pp 233–257 Venema K, van den Abbeele P (2013) Experimental models of the gut microbiome. Best Pract Res

Clin Gastroenterol 27:115–126

Venema K, van Nuenen HMC, Smeets-Peeters M, Minekus M, Havenaar R (2000) TNO's in vitro large intestinal model: an excellent screening tool for functional food and pharmaceutical research. Ernährung/Nutrition 24:558–564

Venema K, van Nuenen MHMC, van den Heuvel EG, Pool W, van der Vossen JMBM (2003) The effect of lactulose on the composition of the intestinal microbiota and short-chain fatty acid production in human volunteers and a computer-controlled model of the proximal large intestine. Microb Ecol Health Dis 15:94–105

< Prev   CONTENTS   Next >