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
- Trypsinize at subconfluency; if needed, split again:
- 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).
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