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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. Changing the media in a flask is slightly different than changing from a transwell monolayer. Here, the procedure for a monolayer will be outlined.

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

Procedure

1. Pre-warm an alliquot of DMEM complete cell culture media.
2. Aspirate media from the outer well and then the inner well using a clean pipette tip and the vacuum pipette (tips in the metal container). When aspirating from the inner well, place the tip of the pipette at an angle against the side wall and carefully lower as to not disturb the monolayer.
3. Take fresh DMEM from the media alliquot and pipette 500μL into the outer well and 100-200μL into the inner well.
4. Observe the cells in the microscope and place back in the incubator.

Quality Control

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

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