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Trypsin Cell Recovery from the Surface of Hydrogels


Introduction

This protocol is for releasing cells from the surface of HyStem®-C and HyStem®-HP hydrogels.

 

Required Materials
  • HyStem-C or HyStem-HP Hydrogels
  • Trypsin

 

Notes
  • Other products (i.e., Accutase, Detachin, TrypLE) that are gentler than trypsin and have greater cell viability can also be used. However, they may also degrade the hydrogel so that recovered cells carry some hydrogel particles with them. If this occurs, then use a 10X collagenase/hyaluronidase solution (StemCell Technologies Cat # 07912) to digest the remaining hydrogel.

Steps
  • 1. Aspirate off media.
     
  • 2. Wash the hydrogel surface with PBS.
     
  • 3. Add trypsin to the hydrogel surface.
     
  • 4. Incubate at room temperature until the cells begin to detach.
     
  • 5. Gently tap the plate to loosen the cells.
     
  • 6. Add media with serum to the hydrogel surface.
     
  • 7. Pipette up and down to get a uniform cell suspension.
     
  • 8. Transfer cells to a culture tube.
     
  • 9. Spin down cells, remove supernatant, and replace with fresh media.

    Note:
     Cell viability will be similar to that of cells grown on plastic and detached with trypsin.



 

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