Decellularizing Mammary Fat Pads: Deriving Extracellular Matrix Hydrogels from Mammary Fat Pads

First, place mammary fat pads or MFPs in a dish with Trypsin-EDTA solution and incubate for the desired duration to break the cell matrix adhesions. Wash the MFPs with deionized water and strain through a strainer. Place tissues in a beaker with a stir bar and add Triton X-100. Stir to lyse cells without affecting the growth factors in the tissue's extracellular matrix.

Strain the MFPs and rinse with deionized water. Transfer tissues back to the beaker, and add deoxycholic acid to lyse the remaining cells. Next, strain the MFPs and place in a beaker containing deionized water with penicillin-streptomycin to prevent contamination. Incubate the beaker at 4 degrees Celsius overnight.

Strain MFPs and place them into a beaker containing ethanol and peracetic acid. The peracetic acid increases the stiffness of the tissue's extracellular matrix. Stir for some time and repeat the straining step. Wash the MFPs with PBS and then with deionized water. Transfer the strained MFPs to a beaker containing propane-1-ol.

Stir for some time, strain the contents, and wash MFPs with deionized water. Dry the MFPs and store at minus 80 degrees Celsius. In the following protocol, we will perform the decel