Patch-clamp recording of Müller glial cells after cryopreservation.
Human and other primate retinal Müller cells display dominating K(+) currents as well as other membrane conductances that may change in cases of retinal pathology. Because the use of human and primate tissue is limited by reasons of availability, a method for long-term storage of these cells is desirable. We describe a cryopreservation method in which isolated Müller cells are stored in liquid nitrogen. After thawing, the cells can be used for patch-clamp experiments immediately, without culturing. We show that the main electrophysiological properties are not altered by this method and that voltage- and ligand-gated currents can be recorded from cryopreserved cells even after 2-years storage.
Biedermann
Bernd
B
Paul-Flechsig-Institut für Hirnforschung, Universität Leipzig, Jahnallee 59, D-04109 Leipzig, Germany.
Wolf
Sebastian
S
Kohen
Leon
L
Wiedemann
Peter
P
Buse
Eberhard
E
Reichenbach
Andreas
A
Pannicke
Thomas
T
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D004533:Egtazic Acid / Q000494:pharmacology; D004594:Electrophysiology; D005615:Freezing; D006801:Humans; D066298:In Vitro Techniques; D008252:Macaca fascicularis; D008564:Membrane Potentials / Q000502:physiology; D008875:Middle Aged; D009457:Neuroglia / Q000502:physiology* / Q000648:ultrastructure; D018408:Patch-Clamp Techniques / Q000295:instrumentation*; D015221:Potassium Channels / Q000502:physiology; D012160:Retina / Q000166:cytology / Q000502:physiology
D015221:Potassium Channels; D004533:Egtazic Acid; D000255:Adenosine Triphosphate