How do I attach cultured mouse Embryoid Bodies (EBs)?

Stem Cell Biology Differentiation
1

Hi

We cultured mouse Embryoid Bodies (EB) for 4 days . Then we were going to let them attach to plate treated by gelatin, but 2 days later, the EBs were still floating in the medium.

Now we want to know this reason why???

Pls help.

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2

Sachin Sharma

You may try the following steps to troubleshoot:

  • Increase gelatin concentration (try 0.2% gelatin) and incubate plates at 37°C for at least 1 hour before seeding.
  • Use alternative coatings like Matrigel, fibronectin, or laminin to enhance adhesion.
  • Extend EB culture to 5–6 days before plating to allow for better differentiation.
  • Reduce medium turbulence by gently pipetting and placing the plate in a static incubator.
  • Check ROCK inhibitors and remove them if used.
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