Comparison between Physical and Enzymatic Harvesting Methods via Flow Cytometry

Mona Zamanian-Azodi; Kobra Rezaei-Tavirani; Maasoumea  Mousavi

Journal of Kermanshah University of Medical Sciences

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Comparison between Physical and Enzymatic Harvesting Methods via Flow Cytometry

Author(s):

Mona Zamanian-Azodi^1,*,

Kobra Rezaei-Tavirani¹,

Maasoumea Mousavi¹

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

Journal of Kermanshah University of Medical Sciences:Vol. 16, issue 8; e77276

Published online:Mar 29, 2013

Article type:Research Article

Received:Dec 03, 2012

Accepted:Feb 19, 2013

How to Cite:Mona Zamanian-AzodiKobra Rezaei-TaviraniMaasoumea MousaviComparison between Physical and Enzymatic Harvesting Methods via Flow Cytometry.J Kermanshah Univ Med Sci.16(8):e77276.

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.

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