Isolation of Adipogenic and Fibro-Inflammatory Stromal Cell Subpopulations from Murine Intra-Abdominal Adipose Depots

The protocol allows investigators to isolate functionally and molecularly distinct subpopulations of perigonadal white adipose tissue stromal cells from mouse models of interest. The advantage of this technique is that all tools and reagents are widely available. Demonstrating the procedure will be Lavanya Vishvanath, a senior laboratory research analyst from Dr.Rana Gupta's laboratory.

After stromal vascular cell isolation from gonadal white adipose tissue, or WAT, resuspend the cell pellet in one milliliter of red blood cell lysis buffer for one to two minute incubation at room temperature. Stop the lysis with 10 milliliters of 2%FBS and PBS and filter the cells through a 40 micron cell strainer into a new 50 milliliter centrifuge tube. Collect the cells by centrifugation and resuspend the pellet in 400 to 800 microliters FBS and PBS, supplemented with FC block.