Cryomilling of Decellularized Tissue: A Technique to Obtain Fine Extracellular Matrix Powder

Use sharp surgical scissors to finely mince the decellularized tissue. To cryomill the processed tissue, fill a 25-milliliter stainless steel milling chamber for a laboratory ball mill system with minced lyophilized ECM and add two 10-millimeter stainless steel milling balls.

Close and completely submerge the loaded milling chamber in liquid nitrogen for 3 minutes. Then, mill the frozen sample at 30 hertz for 3 minutes. Repeat the submersion in liquid nitrogen and the milling until the ECM is milled into a fine powder.