HDP1096 Perfecta3D® 96-Well Hanging Drop Plates Protocol

Cell Culture Media Exchange

We recommend that cell culture media be exchanged every day or every other day to provide enough nutrients for cells and to prevent osmolality shift of the media. The optimal amount of media to be removed and replaced will vary with drop volume, incubator conditions, cell types, and cell densities.

Example media exchange regimens based on 40 μL drop:

  • Every day – either add 4 μL fresh media or remove 10 μL media and replace with 14 μL fresh media.
  • Every other day – either add 10 μL fresh media or remove 14 μL media and replace with 20 μL fresh media.

    **As mentioned above the amount of media to be added or removed will vary with experimental conditions and may require optimization.

Steps:

  1. Pre-warm fresh culture media to 37 °C.
  2. Remove culture media from hanging drops by placing the end of pipette tips at the neck region of the access holes or wells and carefully aspirating media off from the access holes.
  3. Add fresh media back into hanging drops by placing the end of the pipette tips in the neck region of the access holes or wells and slowly dispense fresh media into the access holes.

 

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Materials

     
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