Simple Workflow and Comparison of Media for hPSC-Cardiomyocyte Cryopreservation and Recovery.

Autor: Miller DC; Core Facility Stem Cells, Max-Delbrück-Centrum, Berlin, Germany.; DZHK (German Centre for Cardiovascular Research), partner site Berlin, Berlin, Germany., Genehr C; Core Facility Stem Cells, Max-Delbrück-Centrum, Berlin, Germany., Telugu NS; Core Facility Stem Cells, Max-Delbrück-Centrum, Berlin, Germany., Kurths S; Core Facility Stem Cells, Max-Delbrück-Centrum, Berlin, Germany., Diecke S; Core Facility Stem Cells, Max-Delbrück-Centrum, Berlin, Germany.; DZHK (German Centre for Cardiovascular Research), partner site Berlin, Berlin, Germany.; Berlin Institute of Health (BIH), Berlin, Germany.
Jazyk: angličtina
Zdroj: Current protocols in stem cell biology [Curr Protoc Stem Cell Biol] 2020 Dec; Vol. 55 (1), pp. e125.
DOI: 10.1002/cpsc.125
Abstrakt: Great progress has been made with protocols for the differentiation and functional application of hPSC-cardiomyocytes (hPSC-CMs) in recent years; however, the cryopreservation and recovery of hPSC-CMs still presents challenges and few reports describe in detail the protocols and general workflow. In order to facilitate cryopreservation and recovery of hPSC-CMs for a wide range of applications, we provide detailed information and step-by-step protocols. The protocols are simple and use common reagents. They are comprised of a fast dissociation, cryopreservation using standard equipment, and gentle recovery following thawing. We discuss various features of the protocols, as well as their utilization in the context of common hPSC-CM differentiation and application workflows. Finally, we compare two proprietary and two common in-house formulations of cryopreservation media used for hPSC-CMs, and despite differences in their price and composition find broadly similar recovery rates and cellular function after thawing. © 2019 The Authors. Basic Protocol 1: Dissociation and cryopreservation of hPSC-CMs Basic Protocol 2: Thawing and recovery of cryogenically frozen hPSC-CMs.
(© 2019 The Authors.)
Databáze: MEDLINE
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