title: 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.
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Carver College of Medicine, University of Iowa, 169 Newton Rd., PBDB 4334, Iowa City, IA, 52242, USA.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Carver College of Medicine, University of Iowa, 169 Newton Rd., PBDB 4334, Iowa City, IA, 52242, USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Tong
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Carver College of Medicine, University of Iowa, 169 Newton Rd., PBDB 4334, Iowa City, IA, 52242, USA.
authors.forename: Anil
authors.initials: A
authors.lastname: Mishra
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Carver College of Medicine, University of Iowa, 169 Newton Rd., PBDB 4334, Iowa City, IA, 52242, USA.
authors.forename: Litao
authors.initials: L
authors.lastname: Xie
authors.affiliation: Department of Surgery, University of Iowa, Carver College of Medicine, Iowa City, IA, 52242, USA.
authors.forename: Isaac
authors.initials: I
authors.lastname: Samuel
authors.affiliation: Department of Surgery, University of Iowa, Carver College of Medicine, Iowa City, IA, 52242, USA.
authors.forename: Jessica K
authors.initials: JK
authors.lastname: Smith
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Carver College of Medicine, University of Iowa, 169 Newton Rd., PBDB 4334, Iowa City, IA, 52242, USA. rajan-sah@uiowa.edu.|Fraternal Order of the Eagles Diabetes Research Center, Iowa City, IA, 52242, USA. rajan-sah@uiowa.edu.
authors.forename: Rajan
authors.initials: R
authors.lastname: Sah
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000502:physiology*; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D055724:Electrophysiological Phenomena; D006801:Humans; D051379:Mice; D018408:Patch-Clamp Techniques / Q000379:methods
keywords: Adipocyte isolation; Culture; Differentiation; Patch-clamp; Perforated-patch; Whole-cell patch-clamp