The Use of Short Tandem Repeat Analysis for Cell Line Authentication

Short tandem repeat analysis (STR) is a well-established international method of authentication and genetic stability testing of cell lines (CLs). Therefore, the development and introduction of this method into routine practice of cell banks and cell culture collections is a pressing concern. In add...

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Main Authors: M. D. Khorolsky, I. S. Semenova, E. V. Melnikova, Yu. V. Olefir
Format: Article
Language:Russian
Published: Ministry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products» 2019-12-01
Series:Биопрепараты: Профилактика, диагностика, лечение
Subjects:
Online Access:https://www.biopreparations.ru/jour/article/view/255
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author M. D. Khorolsky
I. S. Semenova
E. V. Melnikova
Yu. V. Olefir
author_facet M. D. Khorolsky
I. S. Semenova
E. V. Melnikova
Yu. V. Olefir
author_sort M. D. Khorolsky
collection DOAJ
description Short tandem repeat analysis (STR) is a well-established international method of authentication and genetic stability testing of cell lines (CLs). Therefore, the development and introduction of this method into routine practice of cell banks and cell culture collections is a pressing concern. In addition, the expansion of the field of cell-line based biomedical cell products (BСPs) necessitates the implementation of STR as a tool of identification testing during quality control. The State Pharmacopoeia of the Russian Federation does not require mandatory use of STR for cell line identification, while other countries have been using this method for cell line quality control for about a decade. The use of identified CLs in medical practice will ensure the efficacy and safety of BCPs. The aim of the study was to assess the possibility of using STR analysis for authentication and genetic stability testing of CLs using U937, WISH, WIL2-S, NK-92, and Jurkat Clone E6-1 CLs as examples. Materials and me­thods: the following human CLs were used in the study: U937 (ECACC), WISH (ATCC), WIL2S (ATCC), NK-92 (ATCC), and Jurkat Clone E6-1 (ATCC). The CL allelic profiles were determined by STR using the COrDIS Plus kit (Gordiz, Russia). The electrophoretic separation was performed using a Genetic Analyzer 3500 Series instrument. The data provided on the websites of the European Collection of Authenticated Cell Cultures and American Type Culture Collection were used to compare the CL profiles. Results: the AuthentiFiler PCR Amplification Kit (Thermo Fisher Scientific, USA) and the GenePrint 10 System (Promega Corporation, USA) intended for CL authentication by STR were compared with the characteristics of the COrDIS plus kit (Gordiz, Russia). The results of the comparison demonstrated that the COrDIS plus kit includes all the loci found in the foreign kits, as well as the loci recommended by the International Cell Line Authentication Committee. The U-937, WIL2S, and NK-92 CLs demonstrated genetic identity with the reference profiles available on the websites of the international collections. The Jurkat Clone E6-1 CL was found to be genetically instable due to the loss of the amelogenin gene. Conclusions: it was demonstrated by the examples of U937, WISH, WIL2-S, NK-92, and Jurkat Clone E6-1 CLs that STR and the COrDIS plus kit could be used for authentication and genetic stability testing. The obtained results suggest the feasibility of using the COrDIS plus kit for the analysis of CLs used in BCPs, for BCP quality control, and biomedical research.
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spelling doaj.art-0aa86ad8e55542a2b8946bcea45f8db02024-03-26T09:10:53ZrusMinistry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products»Биопрепараты: Профилактика, диагностика, лечение2221-996X2619-11562019-12-0119425126010.30895/2221-996X-2019-19-4-251-260191The Use of Short Tandem Repeat Analysis for Cell Line AuthenticationM. D. Khorolsky0I. S. Semenova1E. V. Melnikova2Yu. V. Olefir3Scientific Centre for Expert Evaluation of Medicinal ProductsScientific Centre for Expert Evaluation of Medicinal ProductsScientific Centre for Expert Evaluation of Medicinal ProductsScientific Centre for Expert Evaluation of Medicinal ProductsShort tandem repeat analysis (STR) is a well-established international method of authentication and genetic stability testing of cell lines (CLs). Therefore, the development and introduction of this method into routine practice of cell banks and cell culture collections is a pressing concern. In addition, the expansion of the field of cell-line based biomedical cell products (BСPs) necessitates the implementation of STR as a tool of identification testing during quality control. The State Pharmacopoeia of the Russian Federation does not require mandatory use of STR for cell line identification, while other countries have been using this method for cell line quality control for about a decade. The use of identified CLs in medical practice will ensure the efficacy and safety of BCPs. The aim of the study was to assess the possibility of using STR analysis for authentication and genetic stability testing of CLs using U937, WISH, WIL2-S, NK-92, and Jurkat Clone E6-1 CLs as examples. Materials and me­thods: the following human CLs were used in the study: U937 (ECACC), WISH (ATCC), WIL2S (ATCC), NK-92 (ATCC), and Jurkat Clone E6-1 (ATCC). The CL allelic profiles were determined by STR using the COrDIS Plus kit (Gordiz, Russia). The electrophoretic separation was performed using a Genetic Analyzer 3500 Series instrument. The data provided on the websites of the European Collection of Authenticated Cell Cultures and American Type Culture Collection were used to compare the CL profiles. Results: the AuthentiFiler PCR Amplification Kit (Thermo Fisher Scientific, USA) and the GenePrint 10 System (Promega Corporation, USA) intended for CL authentication by STR were compared with the characteristics of the COrDIS plus kit (Gordiz, Russia). The results of the comparison demonstrated that the COrDIS plus kit includes all the loci found in the foreign kits, as well as the loci recommended by the International Cell Line Authentication Committee. The U-937, WIL2S, and NK-92 CLs demonstrated genetic identity with the reference profiles available on the websites of the international collections. The Jurkat Clone E6-1 CL was found to be genetically instable due to the loss of the amelogenin gene. Conclusions: it was demonstrated by the examples of U937, WISH, WIL2-S, NK-92, and Jurkat Clone E6-1 CLs that STR and the COrDIS plus kit could be used for authentication and genetic stability testing. The obtained results suggest the feasibility of using the COrDIS plus kit for the analysis of CLs used in BCPs, for BCP quality control, and biomedical research.https://www.biopreparations.ru/jour/article/view/255biomedical cell productauthenticity (identification)cell line allelic profilestrcell line
spellingShingle M. D. Khorolsky
I. S. Semenova
E. V. Melnikova
Yu. V. Olefir
The Use of Short Tandem Repeat Analysis for Cell Line Authentication
Биопрепараты: Профилактика, диагностика, лечение
biomedical cell product
authenticity (identification)
cell line allelic profile
str
cell line
title The Use of Short Tandem Repeat Analysis for Cell Line Authentication
title_full The Use of Short Tandem Repeat Analysis for Cell Line Authentication
title_fullStr The Use of Short Tandem Repeat Analysis for Cell Line Authentication
title_full_unstemmed The Use of Short Tandem Repeat Analysis for Cell Line Authentication
title_short The Use of Short Tandem Repeat Analysis for Cell Line Authentication
title_sort use of short tandem repeat analysis for cell line authentication
topic biomedical cell product
authenticity (identification)
cell line allelic profile
str
cell line
url https://www.biopreparations.ru/jour/article/view/255
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