Identification of cross-contaminated animal cells by PCR and isoenzyme analysis.

Cytotechnology
R RamyaV A Srinivasan

Abstract

Animal cell lines have become very popular substrates for the production of vaccines and biopharmaceuticals. Characterization of candidate production cell lines is central to ensure product safety and maintenance of consistency in the manufacture of biologicals. Nested PCR and isoenzyme analysis have been used widely to prove the identity and purity of various cell lines and primary cells individually and also after deliberate cross-contamination. The nested PCR based on the Cytochrome b (Cyt b) gene of mitochondrial DNA (Mt DNA) was found to be more sensitive than isoenzyme analysis in detecting low levels of contaminants (as low as 1%). Interestingly, competition between different co-cultured cell lines has shown in one case that cross-contamination need not always results in a mixed cell population. The nested PCR technique for the Cyt b gene described in this study appears to be a potential replacement for isoenzyme analysis and here we demonstrate the PCR method used is sensitive and reliable for cell line authentication in a simple, rapid and reliable format to help assure the authenticity of cell substrates for the production of safe vaccines and biopharmaceuticals.

References

Jan 1, 1976·American Journal of Hematology·C S StulbergW F Simpson
Jul 4, 1985·Nature·A J JeffreysS L Thein
Apr 24, 1981·Science·W A Nelson-ReesR R Flandermeyer
Feb 3, 1983·Nature·C M O'TooleL M Franks
Feb 1, 1995·In Vitro Cellular & Developmental Biology. Animal·K G SteubeH G Drexler
Mar 1, 1997·Biologicals : Journal of the International Association of Biological Standardization·G N StaceyA Doyle
May 30, 1998·In Vitro Cellular & Developmental Biology. Animal·O Markovic, N Markovic
May 30, 1998·In Vitro Cellular & Developmental Biology. Animal·R W NimsJ W Harbell
Mar 17, 2001·Nature Reviews. Molecular Cell Biology·J R Masters
Feb 3, 2004·In Vitro Cellular & Developmental Biology. Animal·Merry Y LiuAbbas Vafai
Mar 16, 2005·Analytical Biochemistry·María López-AndreoAntonio Puyet
Sep 27, 2005·Alternatives to Laboratory Animals : ATLA·Sandra CoeckeUNKNOWN Second ECVAM Task Force on Good Cell Culture Practice
May 23, 2007·In Vitro Cellular & Developmental Biology. Animal·Kazumi OnoHidekazu Sawada
Nov 13, 2008·Cytotechnology·R Ian Freshney

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Citations

Jun 16, 2015·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Marc-Olivier BaradezDamian Marshall
Oct 17, 2019·Anais Da Academia Brasileira De Ciências·Shirley B DE AraújoMaria Judite B Fernandes
Dec 14, 2018·Nature Communications·Hasan Erbil AbaciAngela M Christiano

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