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A non-BRICHOS surfactant protein c mutation disrupts epithelial cell function and intercellular signaling
Authors:Markus Woischnik  Christiane Sparr  Sun?ana Kern  Tobias Thurm  Andreas Hector  Dominik Hartl  Gerhard Liebisch  Surafel Mulugeta  Michael F Beers  Gerd Schmitz  Matthias Griese
Institution:1. The Biophysical Interdisciplinary Schottenstein Center for the Research and Technology of the Cellome, Bar-Ilan University, Ramat Gan, 52900, Israel
3. WBT Ltd, POB 1516, Ramat Gan, 52115, Israel
4. GeSiM mbH, Bautzner Landstra?e 45, 01454, Gro?erkmannsdorf, Germany
5. Zentrum für gyn?kologische Endokrinologie und Reproduktionsmedizin, Kaiserstrasse 5-7, 66111, Saarbrücken, Germany
2. Main Department for Biophysics and Cryotechnology, Fraunhofer IBMT, Ensheimer Stra?e 48, 66386 St, Ingbert, Germany
6. Professorship for Molecular and Cellular Biotechnology/Nanotechnology, University of Saarland, 66041, Saarbrücken, Germany
Abstract:

Background

Cryopreservation is the only widely applicable method of storing vital cells for nearly unlimited periods of time. Successful cryopreservation is essential for reproductive medicine, stem cell research, cord blood storage and related biomedical areas. The methods currently used to retrieve a specific cell or a group of individual cells with specific biological properties after cryopreservation are quite complicated and inefficient.

Results

The present study suggests a new approach in cryopreservation, utilizing the Individual Cell-based Cryo-Chip (i3C). The i3C is made of materials having appropriate durability for cryopreservation conditions. The core of this approach is an array of picowells, each picowell designed to maintain an individual cell during the severe conditions of the freezing - thawing cycle and accompanying treatments. More than 97% of cells were found to retain their position in the picowells throughout the entire freezing - thawing cycle and medium exchange. Thus the comparison between pre-freezing and post-thawing data can be achieved at an individual cell resolution. The intactness of cells undergoing slow freezing and thawing, while residing in the i3C, was found to be similar to that obtained with micro-vials. However, in a fast freezing protocol, the i3C was found to be far superior.

Conclusions

The results of the present study offer new opportunities for cryopreservation. Using the present methodology, the cryopreservation of individual identifiable cells, and their observation and retrieval, at an individual cell resolution become possible for the first time. This approach facilitates the correlation between cell characteristics before and after the freezing - thawing cycle. Thus, it is expected to significantly enhance current cryopreservation procedures for successful regenerative and reproductive medicine.
Keywords:
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