Caco-2 cell growth

Phillips J, Arena A. Optimization of Caco-2 Cell Growth and Differentiation for Drug Transport Studies. Millipore Corporation Protocol Note PC1060EN00, 2003. 

Kim, E.J., Holthuizen, P.E., Park, H.S., Ha, Y.L., Jung, K.C., and Park, J.H.Y., Trans-10, cis-12-conjugated linoleic acid inhibits Caco-2 colon cancer cell growth, Am. J. Physiol. Gastrointest. Liver Physiol., 283, G357, 2002. 

Kuhfeld MT, Mahraoui L, Rodolosse A et al. (1994) Presence and differential expression of SGLT1, Glutl, Glut2, Glut3 and Glut5 hexose transporter mRNAs in CACO-2 cell clones in relation to cell growth and glucose consumption. Biochem J 298 629-633... 

Complexity is seen in different models. Parathyroid hormone-related protein and Ca have conflicting actions in a nude rat model of humoral hypercalcemia of malignancy. In differentiated Caco cells, there is upregulation of P450 27B1 expression by la,25-dihydroxyvita-min Dj and epidermal growth factor, but down-regulation in less differentiated Caco cell lines . 

Nonintestinal Cell Lines. Madin Darby canine kidney cells (MDCK [50]) differentiate into columnar epithelial cells forming tight Junctions when cultured on semi-permeable membranes they are commonly applied to study cell growth regulation, metabolism, toxicity, and transport at the level of the distal renal tubule epithelia [51]. MDCK cells, like Caco-2 cells, are suitable for molecular permeability screening studies MDCK cells have an advantage over Caco-2 cells in that they do not need 3 weeks in culture before differentiation, but a disadvantage is that they do not express P-gp. 

The other bottleneck for lactic acid production is the operating cost. For example, sterilization is necessary for fermentative production. Hence, microorganisms have an optimal fermentation temperature between 30 2°C (John et al., 2007). Therefore it is difficult to avoid contamination if the medium is not sterilized. Qin et al. (2009) have reported the use of a newly isolated thermophilic strain. Bacillus sp. strain 2 to 6, for the unsterilized fermentative production of L-lactic acid. A high yield (97.3%), productivity (4.37g/L/h), and optical purity of L-lactic acid (99.4%) were obtained in batch and fed-batch open fermentations (Qin et al., 2009). This will help to reduce energy consumption and lower labor costs. Moreover, because of the inhibitory effects of a low pH on cell growth and lactic acid production, CaCOs must be added to maintain a constant pH as a consequence, the regeneration of precipitated calcium lactate is observed (Datta and Henry, 2006). To solve this problem, a sodium lactate-tolerant strain. Bacillus sp. Na-2, was obtained by ion-beam implantation and applied during an L-lactic acid production process (Qin et al., 2010). On the other hand, new processes can be applied to prevent the production of calcium lactate, for example, reverse osmosis, ultrafiltration, electrodialysis, and solvent extraction (Datta and Henry, 2006). 

Hidalgo, I. J. Kato, A. Borchardt, R. T., Binding of epidermal growth factor by human colon carcinoma cell (Caco-2) monolayers, Biochem. Biophys. Res. Commun. 160, 317-324 (1989). 

Baltes S, Nau H, and Lampen A [2004] All-trans retinoic acid enhances differentiation and influences permeability of intestinal Caco-2 cells under serum-free conditions. Dev Growth Differentiation 46 503-514... 

The Caco-2 permeability assay is usually performed in a Transwell device (Figure 18.1). The Transwell contains two compartments a donor and a receiver compartment. The apical donor compartment contains a porous membrane that supports the growth of the Caco-2 monolayer. Caco-2 cells are seeded on the porous membrane. Upon confluency of the cell culture, the compound is added into the donor compartment at a concentration range from one to several hundred micromolar. Samples are collected from the receiver compartment for up to 2 h, then LC-UV or LC-MS methods are used to quantify compound in each sample. The permeability coefficient of the compound is calculated based on the following equation ... 

Hossain, Z., Konishi, M., Hosokawa, M., and Takahashi, K. (2006). Effect of polyunsaturated fatty acid-enriched phosphatidylcholine and phosphatidylserine on butyrate-induced growth inhibition, differentiation and apoptosis in Caco-2 cells. Cell Biochem. Fund. 24, 159-165. 

E Nano, J.L., Nobili, C., Girard-Pipau, F., and Rampal, P., Effects of fatty acids on the growth of Caco-2 cells, Prostaglandins Leukot. Essent. Fatty Acids, 69, 207, 2003. 

L-a-methyldopa a substrate for the amino acid transporter. In Caco-2 cells, the active transport of this dmg by the amino acid transporter was seven times higher than transport by passive diffusion. Its absorption may be further increased by upregulating the amino acid transporter, as has been observed in the 20-70% stimulation of carrier-mediated amino acid transport by treatment of 0.2 mg/kg growth hormone. 

In summary, CACO-2 cells express many enzymes characteristic for intestinal cells involved in drag metabolism. The major drug metabolising enzyme, CYP 3A, is active only in selected clones pointing to polyclonal origin of CACO-2 and the necessity to characterize CACO-2 cells extensively for growth and experimental conditions used for given experiments. 

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