How long does it take for Caco-2 cells to adhere?

How long does it take for Caco-2 cells to adhere?

All Answers (2) 1. when Caco-2 cells are subcultured,it takes about 48h to attach substrate.It may takes more time for thawing cells.

How do Caco-2 cells grow?

The cells should be kept at 37 °C in a humidified atmosphere containing 5 % CO2. 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.

What is the Caco-2 cell line?

Caco-2 is an immortalized cell line of human colorectal adenocarcinoma cells. It is primarily used as a model of the intestinal epithelial barrier. In culture, Caco-2 cells spontaneously differentiate into a heterogeneous mixture of intestinal epithelial cells.

What does Caco-2 stand for?

Freebase. Caco-2. The Caco-2 cell line is a continuous line of heterogeneous human epithelial colorectal adenocarcinoma cells, developed by the Sloan-Kettering Institute for Cancer Research through research conducted by Dr. Jorgen Fogh.

Why are Caco-2 cells used?

Caco-2 cells are a human colon epithelial cancer cell line used as a model of human intestinal absorption of drugs and other compounds. When cultured as a monolayer, Caco-2 cells differentiate to form tight junctions between cells to serve as a model of paracellular movement of compounds across the monolayer.

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) …

How do you freeze Caco-2 cells?

Use the freezing media by Gibco. when yoy add freezing media to cryo tube (1ml), store it for 30 min in freezer, after that 24 hrs at -80°, then store in liquid nitrogen. Hope it will help. Remeber also to store your cells at an early number of passages.

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.

Which compound is represented by Caco-2?

Calcium–cobalt
Calcium–cobalt (1/2)

PubChem CID 71358134
Structure Find Similar Structures
Molecular Formula CaCo2
Synonyms Calcium–cobalt (1/2) DTXSID00782157 221642-44-0
Molecular Weight 157.94

What are calu 3 cells?

Calu-3 cells are a submucosal gland cell line, which was generated from a bronchial adenocarcinoma. Under air–liquid interface conditions Calu-3 cells grow in monolayers, producing mucus and cilia-like microvilli. Because of the presence of TJs, Calu-3 cells are often used for transport studies.

How long does it take Caco cells to double?

As Phyllis mentioned, make sure you are following ATCC recommendations for its culture. IMPORTANT: please note that ATCC reports a doubling time for CACO cells of about 62 hours. This is three times higher when compare to the reported doubling time, for example, of a lung carcinoma cell line as A549 (22 hours).

What should be the viable cell count for Caco-2?

It is important with Caco-2 not to let them reach confluence before subculture. If they grow to confluence they adhere very tightly, are difficult to disaggregate, and do not plate well. Make sure you know the number of viable cells that you are plating after subculture. 2x10e4 or 5x10e4/ml should be about right.

How often should I Change my Caco-2 medium?

Important points for Caco-2 are, never let them become confluent. If they did in the past and frozen like that, they may have been differentiated and grow slowly. 2nd point is, do not use them after passage 50. We are normally changing their medium every other day.

Can you grow caco2-bbe cells in the lab?

I routinely grow Caco2 and Caco2-Bbe cells in the lab with DMEM 10%FBS and they grow fine. It’s important to be sure if you have no mycoplasma. Plus always try to be thorough when you trypsinize. If they are too clumpy when you pass them they will not attach and will die.

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