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

Published: April 30, 2023

Abstract

Source: Galmozzi, A. et al. Isolation and Differentiation of Primary White and Brown Preadipocytes from Newborn Mice. J. Vis. Exp. (2021)

In this video, we demonstrate the surgical procedure to isolate the subcutaneous white adipose tissue and interscapular brown adipose tissue from a newborn mouse.

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) 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 m…

Representative 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…

Divulgaciones

The authors have nothing to disclose.

Materials

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

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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. J. Vis. Exp. (Pending Publication), e20802, doi: (2023).

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