Isolation of sweat glands from mice

Isolation of sweat glands from mice

Materials

• Mouse; C57BL/6NCr, aged 8~12 weeks.
• Culture medium; DMEM (Wako 044-29765) supplemented with 10% fetal bovine serum, penicillin-streptomycin (Gibco 15140122), and GlutaMAX (Thermofisher 35050-061).
• 0.25 mg/mL liberase TM; Add 200 mL of Liberase TM (Roche 5401119001, stock solution 2.5 mg/mL) to 1.8 mL culture medium, prewarmed at 37˚C.

Procedure

• Under deep anesthesia, mouse finger pads were dissected with microscissors and fine-tip forceps under a stereo microscope. The pieces of skin were kept in iced-cold PBS and then minced using a razor blade.
• The dissected skin tissues were collected by brief centrifugation.
• Then, the dissected skin tissues were resuspended in liberase TM (0.25 mg/mL) and digested with periodic mix for 45 minutes at 37˚C. 
• The digested tissues were filtered using a 40 μm-cell strainer, and the isolated sweat glands retained on the filter were collected to be utilized for the subsequent experiments.