Related Experiment Videos
Patch-clamp recording of Müller glial cells after cryopreservation
Bernd Biedermann1, Sebastian Wolf, Leon Kohen
1Paul-Flechsig-Institut für Hirnforschung, Universität Leipzig, Jahnallee 59, D-04109 Leipzig, Germany.
Abstract:
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.
Insights
Cryopreservation allows long-term storage of primate Müller cells for electrophysiology. These retinal cells retain key electrical properties after thawing, even following two years of storage in liquid nitrogen.
Area of Science:
- Neuroscience
- Cell Biology
- Ophthalmology
Background:
- Primate retinal Müller cells are crucial for retinal function.
- These cells exhibit significant potassium (K+) currents and other membrane conductances.
- Retinal pathology can alter Müller cell electrophysiological properties.
Purpose of the Study:
- To develop a method for long-term storage of primate Müller cells.
- To enable immediate use of cryopreserved Müller cells for patch-clamp experiments without culturing.
- To assess the impact of cryopreservation on the electrophysiological properties of Müller cells.
Main Methods:
- Isolated Müller cells were cryopreserved in liquid nitrogen.
- Thawed cells were directly used for patch-clamp experiments.
- Electrophysiological properties, including voltage- and ligand-gated currents, were recorded.
Main Results:
- Cryopreservation did not alter the main electrophysiological properties of Müller cells.
- Functional voltage- and ligand-gated currents were successfully recorded from cryopreserved cells.
- These properties were maintained even after two years of storage in liquid nitrogen.
Conclusions:
- Cryopreservation is a viable method for long-term storage of primate Müller cells.
- Cryopreserved Müller cells can be immediately used for electrophysiological studies.
- This method preserves essential cellular functions for research, overcoming tissue availability limitations.