The individual-cell-based cryo-chip for the cryopreservation, manipulation and observation of spatially identifiable cells. II: Functional activity of cryopreserved cells

BMC Cell Biology

Methodology Article

Steffen Howitz¹  Friederike Ehrhart²  Ina Meiser²  Heiko Zimmermann³  Yaniv Namer⁴  Elena Afrimzon⁴  Naomi Zurgil⁴  Maria Sobolev⁴  Mordechai Deutsch⁴  Yana Shafran⁴  Sergei Moshkov⁴  Assaf Deutsch⁵  Martin Greuner⁶  Michael Thaele⁶ 

[1] GeSiM mbH, Bautzner Landstraβe 45, 01454, Groβerkmannsdorf, Germany;Main Department for Biophysics and Cryotechnology, Fraunhofer IBMT, Ensheimer Straβe 48, 66386, St. Ingbert, Germany;Main Department for Biophysics and Cryotechnology, Fraunhofer IBMT, Ensheimer Straβe 48, 66386, St. Ingbert, Germany;Professorship for Molecular and Cellular Biotechnology/Nanotechnology, University of Saarland, 66041, Saarbrücken, Germany;The Biophysical Interdisciplinary Schottenstein Center for the Research and Technology of the Cellome, Bar-Ilan University, 52900, Ramat Gan, Israel;WBT Ltd., POB 1516, 52115, Ramat Gan, Israel;Zentrum für gynäkologische Endokrinologie und Reproduktionsmedizin, Kaiserstrasse 5-7, 66111, Saarbrücken, Germany;

关键词: Propidium Iodide;    Annexin Versus;    U937 Cell;    Mitochondrial Membrane Potential;    Cryopreserved Cell;   

DOI  :  10.1186/1471-2121-11-83

 received in 2010-05-02, accepted in 2010-10-25,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundThe cryopreservation and thawing processes are known to induce many deleterious effects in cells and might be detrimental to several cell types. There is an inherent variability in cellular responses among cell types and within individual cells of a given population with regard to their ability to endure the freezing and thawing process. The aim of this study was to evaluate the fate of cryopreserved cells within an optical cryo apparatus, the individual-cell-based cryo-chip (i3C), by monitoring several basic cellular functional activities at the resolution of individual cells.ResultsIn the present study, U937 cells underwent the freezing and thawing cycle in the i3C device. Then a panel of vital tests was performed, including the number of dead cells (PI staining), apoptotic rate (Annexin V staining), mitochondrial membrane potential (TMRM staining), cytoplasm membrane integrity and intracellular metabolism (FDA staining), as well as post-thawing cell proliferation assays. Cells that underwent the freezing - thawing cycle in i3C devices exhibited the same functional activity as control cells. Moreover, the combination of the multi-parametric analysis at a single cell resolution and the optical and biological features of the device enable an accurate determination of the functional status of individual cells and subsequent retrieval and utilization of the most valuable cells.ConclusionsThe means and methodologies described here enable the freezing and thawing of spatially identifiable cells, as well as the efficient detection of viable, specific, highly biologically active cells for future applications.

【 授权许可】

Unknown   
© Afrimzon et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.