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When to Change

Use this page when culturing CaCo2 cells, to know when to change the medium.

Purpose

Changing the medium is important for maintaining the growth and health of a cell culture. The medium needs to be changed on a routine basis and it is essential to do it at the right time and with the right procedure as to not contaminate the cell culture.

Safety

• Complete work in the BSC.
• Spray down all materials brought into the BSC with 70% Ethanol.
• Try not to touch the sides of the flask opening with pipette tips.
• Do not touch the rims of the flasks or tubes to work with.

Materials

• Cell culture flask with seeded cells
• Motorized pipette
• Glass pipette tips with vacuum aspiration
• Complete DMEM media (Caco-2 media recipe)

Procedure

1. Preheat an alliquot of complete DMEM media in the 37°C water bath for 10-15 minutes, until fully warm. Retrieve the cell culture flask from the incubator.
2. In a clean BSC cleaned with 70% ethanol, use a glass pipette from the metal container and change the old medium from the flask. Do not make contact with the rim of the flask and try not to make contact with the cells adhered to the base of the flask.
3. Add 12-15mL of new complete DMEM media to the flask using a motorized pipette, ensuring to cover the whole surface area of the flask by tilting and sliding the flask along the surface of the table in a figure-8 motion.
4. Return the flask into the incubator and the DMEM media to the 4° fridge.

Quality Control

• Before changing the media, it is best to check the cell culture flask to see the status of the cell culture. Check tight junction formation and confluence.

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