How should preadipocytes be seeded for optimal differentiation?

Recommended procedure

• Thaw the vial according to the PromoCell protocol.

• Seed part of the cells (5,000 cells/cm²) directly into multiwell plates intended for differentiation.

  • Culture in Preadipocyte Growth Medium until 100% confluency is reached.

  • Then induce differentiation following the recommended protocol.

• Seed the remaining cells (5,000 cells/cm²) into a tissue culture dish for expansion in Preadipocyte Growth Medium.   

  • Trypsinize at subconfluency; if needed, split again:

    • one part for differentiation assays

    • one part to maintain undifferentiated HWP cultures

Important note

• Differentiation capacity may decline after 1-2 passages in vitro.

• Therefore, perform differentiation experiments at early passages.

• Cells at higher passages can still be used for experiments requiring undifferentiated preadipocytes (e.g. proliferation studies).