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How to recover cells from QGel MT 3D Matrix for subculture ?

Céline Gandar
posted this on October 11, 2010 16:49

Please find below an example on how to recover cells grown in QGel™ MT 3D Matrices for subculture:

 

 

How to recover cells from QGel™ MT 3D Matrix for subculture


ABOUT THIS PROTOCOL: This is an example of how to extract cells from QGel™ MT 3D Matrix for subculture. Note that this is only a guideline; reaction time, type and concentrations of proteases should be optimized depending on specific experimental conditions.

 

Suggested chemicals/solutions/kits:

  • TrypLE™ 1X (Invitrogen™)

 

Brief procedure description: 

  1. Transfer desired gel samples (e.g. with a volume of approx. 30-40µL) containing the encapsulated live cells in a new well plate (under cell culture hood) and wash them once with PBS for 20 minutes.
  2. Add 500µL trypLE 1X solution to the gels in each well and replace the trypLE 1X solution every 15 minutes until the gels are degraded.

Note: To accelerate the gel digestion process, gels may be cut into pieces using a sterile razor blade/scalpel and processed as described above.

  1. At each trypLE solution change, collect the solution/cell suspension and mix it 1:1 with fresh cell culture media containing serum to inactivate trypsin.
  2. Check degradation of the hydrogels under the microscope. If digestion is incomplete gel pieces can be seen floating in the solution.
  3. Note: Digestion process may last 60-90 min for the entire gel (approx. gel volume 30-40µL) and 30-40 min if each gel was cut into pieces. Centrifuge the collected gel digestion solution/cell suspension and aspirate the supernatant.
  4. The cell pellets can be resuspended in fresh media and subcultured.