Isolation and Patch-Clamp of Primary Adipocytes

Abstract

The patch-clamp technique allows for the study of ion channel activity in the native adipocyte environment to better understand the contributions of ion channels to adipocyte signaling. Here, we describe methods for isolating primary mature adipocytes from both mouse and human white adipose tissues (subcutaneous and visceral). From the same preparation, we describe how to culture and differentiate preadipocytes isolated from the stromal vascular fraction. We then describe in detail patch-clamp methods, including both whole-cell and perforated-patch configurations.

Keywords: Adipocyte isolation; Culture; Differentiation; Patch-clamp; Perforated-patch; Whole-cell patch-clamp.