NOTICE: You are viewing a page of the openwetware wiki. Our "dewikify" feature makes a wiki page appear as a normal web page. On September 1st 2017, this feature will GO AWAY and this URL will redirect to the source URL on our wiki. We're sorry for the inconvenience.



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


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



  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)


Used in Stanford for Tissue Engineering Lab Course (ME385B)



or instead, discuss this protocol.