Cell Harvesting Methods Affect Cellular Integrity of Adherent Cells During Apoptosis Detection

Publication type:
Journal article
Metadata:
Autoren
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000451742800009&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.21873/anticanres.13034
eISSN
1791-7530
Externe Identifier
  • Clarivate Analytics Document Solution ID: HC3ZR
  • PubMed Identifier: 30504375
ISSN
0250-7005
Ausgabe der Veröffentlichung
12
Zeitschrift
ANTICANCER RESEARCH
Schlüsselwörter
  • Cell death
  • cellular technique
  • flow cytometry
  • cell harvesting method
  • adherent cells
  • apoptosis
Paginierung
6669 - 6672
Datum der Veröffentlichung
2018
Status
Published
Titel
Cell Harvesting Methods Affect Cellular Integrity of Adherent Cells During Apoptosis Detection
Sub types
  • Article
Ausgabe der Zeitschrift
38

Data source: Web of Science (Lite)

Other metadata sources:
Autoren
DOI
10.21873/anticanres.13034
eISSN
1791-7530
ISSN
0250-7005
Ausgabe der Veröffentlichung
12
Zeitschrift
Anticancer Research
Sprache
en
Online publication date
2018
Paginierung
6669 - 6672
Datum der Veröffentlichung
2018
Status
Published
Herausgeber
Anticancer Research USA Inc.
Herausgeber URL
http://dx.doi.org/10.21873/anticanres.13034
Datum der Datenerfassung
2022
Titel
Cell Harvesting Methods Affect Cellular Integrity of Adherent Cells During Apoptosis Detection
Ausgabe der Zeitschrift
38

Data source: Crossref

Abstract
<h4>Background/aim</h4>Annexin V and propidium iodide (PI) dual staining is commonly applied in bioscience as a method to detect apoptosis. However, excessive handling of adherent cells may interfere with the integrity of plasma membrane and hence impede the accuracy of this method. Here, we exploited PI uptake as an indicator of cell integrity and investigated how cell harvesting methods and solutions involved in common apoptosis detection techniques affected measurement results.<h4>Materials and methods</h4>Different cell harvesting techniques, staining with PI and flow cytometry were performed.<h4>Results</h4>Non-fixed scrapped cells revealed significantly higher fractions of PI-positive staining compared to non-fixed trypsinized cells. In the case of harvesting cells by scrapping, samples stained in binding buffer (68.30±3.55%) showed consistently higher PI-positive staining than samples stained in PBS (36.37±5.90%) in a significant manner (p=0.015).<h4>Conclusion</h4>Enzymatic harvesting using 0.25% trypsin instead of mechanical harvesting by rubber scraper caused less damage of cell integrity. Furthermore, the binding buffer used in the apoptosis detection protocol aggravated the existing plasma membrane damage caused by the rubber scraper.
Addresses
  • Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Mainz, Germany.
Autoren
DOI
10.21873/anticanres.13034
eISSN
1791-7530
Externe Identifier
  • PubMed Identifier: 30504375
Open access
false
ISSN
0250-7005
Ausgabe der Veröffentlichung
12
Zeitschrift
Anticancer research
Schlüsselwörter
  • Cell Line, Tumor
  • Humans
  • Propidium
  • Trypsin
  • Annexin A5
  • Biological Assay
  • Flow Cytometry
  • Cell Culture Techniques
  • Cell Separation
  • Staining and Labeling
  • Cell Adhesion
  • Apoptosis
Sprache
eng
Medium
Print
Paginierung
6669 - 6672
Datum der Veröffentlichung
2018
Status
Published
Datum der Datenerfassung
2018
Titel
Cell Harvesting Methods Affect Cellular Integrity of Adherent Cells During Apoptosis Detection.
Sub types
  • Journal Article
Ausgabe der Zeitschrift
38

Data source: Europe PubMed Central

Abstract
BACKGROUND/AIM: Annexin V and propidium iodide (PI) dual staining is commonly applied in bioscience as a method to detect apoptosis. However, excessive handling of adherent cells may interfere with the integrity of plasma membrane and hence impede the accuracy of this method. Here, we exploited PI uptake as an indicator of cell integrity and investigated how cell harvesting methods and solutions involved in common apoptosis detection techniques affected measurement results. MATERIALS AND METHODS: Different cell harvesting techniques, staining with PI and flow cytometry were performed. RESULTS: Non-fixed scrapped cells revealed significantly higher fractions of PI-positive staining compared to non-fixed trypsinized cells. In the case of harvesting cells by scrapping, samples stained in binding buffer (68.30±3.55%) showed consistently higher PI-positive staining than samples stained in PBS (36.37±5.90%) in a significant manner (p=0.015). CONCLUSION: Enzymatic harvesting using 0.25% trypsin instead of mechanical harvesting by rubber scraper caused less damage of cell integrity. Furthermore, the binding buffer used in the apoptosis detection protocol aggravated the existing plasma membrane damage caused by the rubber scraper.
Date of acceptance
2018
Autoren
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/30504375
DOI
10.21873/anticanres.13034
eISSN
1791-7530
Ausgabe der Veröffentlichung
12
Zeitschrift
Anticancer Res
Schlüsselwörter
  • Cell death
  • adherent cells
  • apoptosis
  • cell harvesting method
  • cellular technique
  • flow cytometry
  • Annexin A5
  • Apoptosis
  • Biological Assay
  • Cell Adhesion
  • Cell Culture Techniques
  • Cell Line, Tumor
  • Cell Separation
  • Flow Cytometry
  • Humans
  • Propidium
  • Staining and Labeling
  • Trypsin
Sprache
eng
Country
Greece
Paginierung
6669 - 6672
PII
38/12/6669
Datum der Veröffentlichung
2018
Status
Published
Datum, an dem der Datensatz öffentlich gemacht wurde
2019
Titel
Cell Harvesting Methods Affect Cellular Integrity of Adherent Cells During Apoptosis Detection.
Sub types
  • Journal Article
Ausgabe der Zeitschrift
38

Data source: PubMed

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