Caco-2 cells have special features: they develop into small intestinal cells with tight junctions and microvilli, expressing the drug metabolizing enzymes involved in uptake of the agents crucial in drug metabolism [22,23].
How do you differentiate Caco-2 cells?
Standard methods used to differentiate Caco-2 cells involve seeding them on culture inserts fitted with polycarbonate filters, and allowing spontaneous differentiation to proceed for two to three weeks in culture medium containing 10% or 20% foetal bovine serum (FBS) in both the apical (AP) and basolateral (BL)
What is a good Caco-2 permeability?
Most drugs are not especially permeable through Caco-2 cells, with the median Papp value being some 16 ⋅ 10−6 cm s−1. This value is considerably lower than those (1,310 and 230 ⋅ 10−6 cm s−1) recently used in some simulations that purported to show that Papp values were too great to be transporter-mediated only.17 Nov 2015
What is A549 cell line?
A549 cells are lung carcinoma epithelial cells that constitute a cell line. A549 is one of the commonly used cell lines for a wide range of research applications.
How do you split Caco-2 cells?
For propagation in culture flasks, Caco-2 cells are seeded in a concentration of 105 cells/cm2. Medium should be changed every 3 days. At 80 % confluence, typically after 4–5 days, the cells are split 1:10 before further cultivation. Trypsinize Caco-2 cells by first rinsing with PBS (15 ml pr.
What is the Caco-2 cell line?
Caco-2 [Caco2] are epithelial cells isolated from colon tissue derived from a 72-year-old, White, male with colorectal adenocarcinoma. This cell line is a suitable transfection host and has applications in cancer and toxicology research.
Where are Caco-2 cells?
The Caco-2 cell line is originally derived from a colon carcinoma. However, one of its most advantageous properties is its ability to spontaneously differentiate into a monolayer of cells with many properties typical of absorptive enterocytes with brush border layer as found in the small intestine.
Why are Caco-2 cells used?
The human epithelial cell line Caco-2 has been widely used as a model of the intestinal epithelial barrier. 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.
How do Caco-2 cells grow?
During growth, Caco-2 cells go through processes of proliferation, confluency, and differentiation. When grown under standard conditions on semipermeable membranes, fully differentiated Caco-2 cells are very similar to normal enterocytes with regards to their morphological characteristics.
What is a major limitation to the Caco-2 monolayer permeability assay?
in 2007 regarding the use of Caco-2 monolayers in permeability assays [15]. The major limitation of this reference protocol is its very low throughput due to the long culture period (at least 21 days), although this has been shown to be required to obtain a fully differentiated and confluent monolayer [16,17].26 Sept 2021
What is Caco permeability?
Cyprotex's Caco-2 permeability assay uses an established method that measures the rate of flux of a compound across polarised Caco-2 cell monolayers and from which the data generated can be used to predict in vivo absorption of drugs. The Caco-2 cell line is derived from a human colon carcinoma.
How long does it take for Caco-2 cells to adhere?
Caco-2 cells were either differentiated for 21 days as described above or seeded on permeable supports (12-well) at a seeding density of 100,000 cells/well which corresponds to 8.9 × 104 cells/cm2 and allowed to adhere for 24 h (referred to as undifferentiated).21 Jun 2017