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Contents

Overview

This protocol will induce adipocytic differentiation in 3T3 Fibroblasts that can be visualized by Oil Red O staining.

Materials

450 ml DMEM (Gibco #11995065) + 50 ml Calf Serum (Gibco #16010159)

Solutions


Procedure

  1. Day 1
    1. Heat complete medium, trypsin/EDTA, and PBS to 37C in water bath
    2. Subculture 3T3 cells with the following concentrations: (25,000cells/cm2)
      • 6 well plate: 2.27x105cells/well in 6ml media
      • tissue culture dish: 1.42x106cells/dish in 10ml media
    3. Place cells in incubator for 1-2hr
    4. Replace growth media with Adipogenic Media
  2. Day 3
    • Replace Adipogenic Media
  3. Day 4
    • Change from Adipogenic Media to Growth Media + Insulin (adipocyte maintenance)
  4. Day 6
    • Replace Growth Media + Insulin (adipocyte maintenance)


Notes

Used in Stanford for Tissue Engineering Lab Course (ME385B)

References

Contact


or instead, discuss this protocol.