The expression of the brush borderassociated hydrolase sucraseisomaltase was shown to increase from early to late passages of caco 2 cells, concomitant with a decrease in the rates of glucose consumption. Compozr admetox cell lines caco2 bcrp knockout cell line 96 well assay ready plate catalog number mtox1002p96 store at room temperature technical bulletin product description compozr zinc finger nuclease zfn technology is a fast and reliable way to manipulate the genome in a. In view of these observed similarities with iron absorption studies in humans, we conclude that the caco2 cell culture sys. When cultured as a monolayer, caco2 cells differentiate to form tight junctions between cells to serve as a model of paracellular movement of. Elucidation of the impact of cell culture conditions of. Human intestinal cell line caco 2t antimicrobial agents and. The model mimics processes such as transcellular transport, paracellular transport, and some aspects of efflux and active transport. Zinc transport and metallothionein secretion in the.
Culturing caco 2 cells with 10mm butyric acid and 15% fbs for 4 days enhanced the basal levels of. The caco 2 cell line as a model of the intestinal barrier. Culturerelated conditions, as well as the different caco2 cell lines utilized in different laboratories, often make it extremely difficult to compare results in the. One such epithelial cell line that has been widely used as a model system of intestinal permeability is the caco 2 cell line. Feed cells by changing media approximately once every two days. The functional differentiation of these cells occurs in. Pdf the caco2 cell line as a model of the intestinal barrier. Navicyte scientific holds the exclusive commercial distribution rights to the caco2 cell line deposited by the. For full access to this pdf, sign in to an existing account, or purchase an annual subscription. According to biopharmaceutics classification system. Various in vitro methods are listed in united states fda guidelines, accep table to evaluate the permeability of a drug substance, includes monolayer of suitable epithelial cells. Caco2 cell cultures in the assessment of intestinal. The caco2 cell line is originally derived from a colon carcinoma. The caco2 cell line is a continuous line of heterogeneous human epithelial colorectal adenocarcinoma cells, developed by the sloankettering institute.
The parental cell line, originally obtained from a human colon adenocarcinoma, undergoes in culture a process of spontaneous differentiation that leads to the formation of a monolayer of cells, expressing several morphological and functional characteristics of. The mechanism of intestinal secretion of the difluorinated quinolone sparfloxacin was investigated with the epithelial cell line caco 2 and was compared to that of the pglycoprotein pgp substrate vinblastine. Caco 2 cells human colorectal adenocarcinoma, atcc. The purpose of this study was to determine whether caco2 cells undergo similar differentiation when grown on transwell polycar bonate membranes, and to. A polymerbased transfection caco2 kit for colon adenocarcinoma cells yields at least 94% transfection efficiency and is a helpful tool for gene expression studies as well as for assessing cellular functionality. Altogen biosystems provides efficient biodegradable transfection reagent kits for the caco2 colon cancer cell line. Development of a caco2 cell line carrying the human. The human colon carcinoma cell line caco 2, when seeded on.
Given the above background, in this study, we sought to develop a new caco 2 cell line that carries a human small. Attempts to either culture intestinal epithelial cells or to establish cell lines derived from the enterocyte have not been successful 5,6. Caco2 is a human colorectal adenocarcinoma cell line isolated in the late 1970s from colon carcinoma tissue1. These clones show considerable and inverse differences in the levels of sucrase activities. Methodology 7,8 cell lines used for screening are should be precultivated. Adhesionofetecstrains nolongeroccurredwhenthe caco 2 cells were pretreated with the homologous colonization factor antigen or when the bacterial cells were. Pdf the caco2 cell line as a model of the intestinal. Evaluation of optimized caco2 permeability measurements oksana tsinman, konstantin tsinman,and alex avdeef pion inc. Caco2 cells were grown on polycarbonate filters in a 1. Drugpermeability and transporter assays in caco2 and. The human colon adenocarcinoma caco 2 and madindarby canine kidney epithelial cell lines provide in vitro tools to assess a drugs permeability and transporter interactions during discovery and development. We conclude that caco2 cells grown on collagencoated polycarbonate mem branes should represent a valuable transport model system for the small intestinal. The caco2 cell line as a model of the intestinal barrier.
Caco2 caco2 atcc htb37 homo sapiens colon colorectal. Forms moderately well differentiated adenocarcinomas consistent with colonic primary grade ii, in nude mice. Thawing, propagating and cryopreserving of ncipbcfhtb37 caco 2 page 4 of 25 special growth requirements subculture caco 2 cells at 80 % to 90 % confluence or when cell. The function, protein levels, and the specific gene deletion of these transporters have been confirmed in both the ko and dko cells. Several possible in vitro human cell models are available for this purpose, one of which is the caco 2 cell model, a well characterised cell line. Pdf the human intestinal caco2 cell line has been extensively used over the last twenty years as a model of the intestinal barrier. Caco2 permeability assay caco 2 permeability goal to measure directional caco 2 permeability of test compounds in.
Vigorously resuspend caco 2 cell pellet with 4610 ml media for 1. Because histamine is a modulator of cancer cell proliferation and invasiveness, this study aimed at investigating the effect of lathyrus sativusderived diamine oxidase lsao and its mechanism of action on caco. The parental cell line, originally obtained from a human colon adenocarcinoma, undergoes in culture a process of spontaneous differentiation that leads to the formation of a monolayer of cells, expressing several morphological and functional characteristics of the mature enterocyte. Caco 2 09042001 transmission electron micrograph tem of caco 2 cells mvmicrovilli. The negative variants, which were cured of the plasmid encodingthe adhesivefactor, did notadhere. Different cell line models used in the drug permeability studies, their characteristics has been summarized emphasizing on caco2. Caco 2 cells were grown until fully confluent on inserts in 12. Development of in vitro pharmacokinetic screens using caco.
The human intestinal caco2 cell line has been extensively used over the last twenty years as a model of the intestinal barrier. Responses of the differentiated intestinal epithelial cell. Secretion of sparfloxacin from the human intestinal caco2. This process was enhanced following exposure from the apical compartment to increasing concentrations of the metal. Lathyrus sativus diamine oxidase counteracts histamine. The caco2 cell membrane permeability of three series of peptides and endothelin antagonists could be predicted by a theoretical model that considered both the polar psad and nonpolar npsa d parts of the dynamic molecular surface area of the molecules. The caco2 cell line in studies of drug metabolism and efflux. Cells were grown in plastic cell culture flasks in dulbeccos modified eagles medium supplemented with 3. Thus, cultivation conditions can be expected to select for the growth of subpopulations of cells resulting in a cellular model system with properties that may differ from the original cell line. Pharmacological effects of cannabinoids on the caco2 cell. Caco2 cells imaged with phase contrast microscopy using 100x magnification. Introduction since many decades, the prediction of oral drug absorption in humans is performed through the use of the caco 2 model 1. The caco2 cell culture system can be used as a model to study. However, the effects of cannabinoids on intestinal permeability have not yet been established.
In this study, the effects of the components in tea on the bioavailability of fluoride were evaluated in the caco. Cell culture is very important approach to investigate various molecular mechanisms and phenotypes in cells that are distinct or separated from whole animal system. Aa305219 est176211 colon carcinoma caco2 cell line ii homo sapiens cdna 5 end similar to. The heat map displays a general higher fold change in the group of selected genes for differentiated caco 2 cells.
Pdf caco2 cells develop morphologic characteristics of normal enterocytes when grown on plastic dishes or nitrocellulose filters. For adherent cell lines, harvest as close to 8090% confluency, as possible. Aa305354 est176335 colon carcinoma caco2 cell line ii homo sapiens cdna 5 end similar to similar to villin 2, mrna sequence. Characterization of the human colon carcinoma cell line caco2. Caco2 cells are continuous lines of heterogenous human epithelial colorectal adenocarcinoma cells. Tea components influencing bioavailability of fluoride and.
In the caco 2 hepatocyte combination model, results from a caco 2 cell permeability. Cac, caco, caco2, caco 2 human caucasian colon adenocarcinoma isolated from a primary colonic tumour in a 72yearold caucasian male using the explant culture technique. Indeed, although cancerous in origin, caco 2 cells are unique in their ability to initiate spontaneous differentiation on reaching confluence under nor mal culture conditions i. Previous work showed that the bioavailability of fluoride in dark tea was lower than naf solution. Transient mosaic patterns of morphological and functional. Comparison of function and relative transporter protein. Heat map representing the fold change of rna levels in the colon adenocarcinoma cell line, caco 2, in both differentiated d and proliferative p states upon interaction with giardia intestinalis gs isolate for 1. Caco 2 cell line was obtained from the european collection of cell cultures ecacc. Adhesion enterotoxigenic escherichia coli to carcinoma. Ornithine decarboxylase and diamine oxidase in human. Aa305219 est176211 colon carcinoma caco2 cell line ii homo sapiens cdna 5 end similar to apolipoprotein b100, mrna sequence.
Remove medium and rinse the adherent cells using pbs without calcium and magnesium 35 ml pbs for t25, 510ml for t75 cell culture flasks. An established model for smallintestinal enterocytes is the caco2 cell line or the caco2 tc7 subclone, derived from a colon adenocarcinoma. Hepcidin attenuates zinc efflux in caco2 cells the. One such epithe lial cell line that has been widely used as a model system of intestinal permea bility is the caco2 cell line. Caco2 cells are a human colon epithelial cancer cell line used as a model of human intestinal absorption of drugs and other compounds. Activation of cannabinoid receptors decreases emesis, inflammation, gastric acid secretion, and intestinal motility. Caco2 bcrp knockout cell line mtox1002p96 technical. When cultured as a monolayer, caco2 cells differentiate to form tight junctions between cells to serve as a model of paracellular movement of compounds across the monolayer. The caco 2 cell line in studies of drug metabolism and efflux sanna siissalo academic dissertation to be presented, with the permission of the faculty of pharmacy of the university of helsinki, for public examination in lecture room 2041, viikki biocenter 2, on 4th april 2009, at 10. Caco 2cell numbera, cellularproteinb, andsucrase activitycatvarioustimesafterconfluence. The endogenous levels of mdr1, bcrp, and mrp2 in the caco 2 wt cells are sufficient to transport substrates. Twentysix clones were isolated from early p29 and late p198 passages of the cell line.
Among these they were able to transport zinc from the apical to the basal compartment. Since most drugs are known to absorb via intestines without using cellular pumps, passive permeability models have came into the limelight. Company identification cls cell lines service gmbh dr. Recently, human intestinal cell lines capable of differentiating in vitro, e. The human epithelial cell line caco2 has been widely used as a model of the intestinal epithelial barrier. Human colon adenocarcinoma cell line, permanent cell line 2. Caco2 cells in this image had been kept in culture for 21 days, allowing them to grow confluent and epithelialize. Material safety data sheet for cell cultures biosafety. We examined in this study whether these yeasts also altered the production of other cytokines and cyclooxygenases coxs by caco 2 cells.
Cytokine responses of intestinal epitheliallike caco2. However, limited information is available indicating the effects of tea components on the fluoride bioavailability. The cells, when cultured on semiporous filters, form confluent monolayers that model the intestinal epithelial barrier for permeability, transporter and druginteraction assays. The aim of the present study is to examine the effects of cannabinoids on intestinal permeability in an in vitro model. Effects of butyrate on intestinal barrier function in a. Differential expression of sucraseisomaltase in clones. Pdf caco2 cell permeability assays to measure drug. Th e generalized culturing and experimental setup for caco2 cell model is as follows.