White and Brown Adipose Depot Collection from Mouse Pup: A Surgical Procedure to Harvest the White Adipose Tissue and Brown Adipose Tissue from Mouse Pup

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Collection and digestion of adipose depots (day 1)

1. Prepare two 1.5 mL tubes for each pup: one for brown adipose tissue (BAT) and one for white adipose tissue (WAT). Add 250 µL of phosphate-buffered saline (PBS) + 200 µL of 2x isolation buffer (123 mM NaCl, 5 mM KCl, 1.3 mM CaCl₂, 5 mM glucose, 100 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), penicillin-streptomycin, and 4% fatty acid-free bovine serum albumin) to each tube. Keep....

Results

Figure 1: Collection and processing of fat pads. (A) Schematic representation of primary white (top) and brown (bottom) adipocyte isolation. (B) Subcutaneous white (top) and brown (bottom) adipose depots. In P0 mice, subcutaneous WAT is almost invisible, but becomes distinguishable on ~day 2 after birth. In contrast, BAT has a distinct dark color even at P0. In o.......

Materials

Name	Company	Catalog Number	Comments
6-well plates	Corning	353046
Fatty Acid-Free BSA	Sigma-Aldrich	A8806
CaCl2	Sigma-Aldrich	C4901
Glucose	Sigma-Aldrich	G7021
HEPES	Sigma-Aldrich	H3375
KCl	Sigma-Aldrich	P9333
NaCl	Sigma-Aldrich	S7653
Pen/Strep	Gibco	15140122
Surgical forceps	ROBOZ Surgical Instrument Co	RS-5158
Surgical Scissors	ROBOZ Surgical Instrument Co	RS-5880
DPBS, no calcium, no magnesium	Gibco	14190144