Comparison between Physical and Enzymatic Harvesting Methods via Flow Cytometry

AUTHORS

Mona Zamanian-Azodi 1 , * , Kobra Rezaei-Tavirani 1 , Maasoumea Mousavi 1

AUTHORS INFORMATION

1 Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

ARTICLE INFORMATION

Journal of Kermanshah University of Medical Sciences: 16 (8); e77276
Published Online: March 29, 2013
Article Type: Research Article
Received: December 03, 2012
Accepted: February 19, 2013

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Abstract

Background: Cell culture is used in laboratory for variety of purposes. In addition, there are different cell harvesting methods. One of these methods is enzymatic method. In this method cells are separated from the plate via using enzyme such as Trypsin. This method, along with side effects of enzyme digestion effect, may affect other parts of cell. To avoid this, via using physical method cells would be separated from the plate. If this method be used it seems that, physical method could possibly damage survival of the cells. In this study, two kinds of damages were compared using Flow Cytometry.   

Methods: MKN45 gastric cancer cells were cultured in RPMI at 37 ºC and 5 %CO2   and after reaching to 80% density, they were separated from the plate using both physical and digestion methods. Then cell status was evaluated and analyzed using flow cytometry method.

Results: Results showed that cell death rate was higher using physical method than enzymatic method. This significant difference was confirmed by microscopic observations.

Conclusion: Since using enzymatic method leads to low cell death rate, it seems that using this method is better than physical method for separation of cells from the plate.

Keywords

Cell Culture Harvesting Trypsin Scraper

© 2013, Journal of Kermanshah University of Medical Sciences. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
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