Preprint Communication Version 1 Preserved in Portico This version is not peer-reviewed

Single Ion Channel Recording in 3D Culture of Stem Cells Using Patch-Clamp Technique

Vladislav I. Chubinskiy-Nadezhdin
* ,
Anastasia V. Sudarikova
ORCID logo ,
Mariia A. Shorokhova
ORCID logo ,
Valeria Y. Vasileva
,
Zuleikha M. Khairullina
,
Yuri A. Negulyaev
Version 1 : Received: 6 April 2022 / Approved: 8 April 2022 / Online: 8 April 2022 (03:29:15 CEST)

A peer-reviewed article of this Preprint also exists.

Chubinskiy-Nadezhdin, V. I.; Sudarikova, A. V.; Shorokhova, M. A.; Vasileva, V. Y.; Khairullina, Z. M.; Negulyaev, Y. A. Single Ion Channel Recording in 3D Culture of Stem Cells Using Patch-Clamp Technique. Biochemical and Biophysical Research Communications, 2022, 619, 22–26. https://doi.org/10.1016/j.bbrc.2022.06.022. Chubinskiy-Nadezhdin, V. I.; Sudarikova, A. V.; Shorokhova, M. A.; Vasileva, V. Y.; Khairullina, Z. M.; Negulyaev, Y. A. Single Ion Channel Recording in 3D Culture of Stem Cells Using Patch-Clamp Technique. Biochemical and Biophysical Research Communications, 2022, 619, 22–26. https://doi.org/10.1016/j.bbrc.2022.06.022.

Abstract

Tri-dimensional (3D) cell aggregates or spheroids are considered to be closer to physiological conditions than traditional 2D cell culture. Mesenchymal stem cells (MSCs) assembling in spheroids have increased the survival of transplanted cells. The regulation of the biological processes that maintain crucial physiological reactions of MSCs is closely related to the functioning of ion channels. The pattern of expression, role and regulatory mechanisms of ion channels could be significantly different in 3D compared to 2D culture, and, thus, needed to be properly analyzed on the level of ionic currents. We developed a specific approach that allowed us to register, for the first time, endogenous ion channels in endometrial MSCs (eMSCs) assembled in spheroids. Particularly, using the single-channel patch-clamp technique, we have recorded the activity of ion channels and observed their functional interplay in mechanosensitive clusters. Our experimental protocol could be applied for identification and studying of ion channels in 3D cell cultures.

Keywords

3D cell culture; patch-clamp; spheroids; single-channel recording; ion channels; mesenchymal stem cells

Subject

Biology and Life Sciences, Cell and Developmental Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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