Justification of Methodological Approaches to Identification Testing of Biomedical Cell Products

Abstract

The manufacturer (developer) has to prepare a specification for each newly developed biomedical cell product (BCP) that has passed the stage of preclinical studies. The specification is included into the registration dossier when applying for marketing authorisation of a BCP. In accordance with the Order of the Ministry of Health of the Russian Federation No. 14n of 19 January 2017 «On approval of the specification format for a biomedical cell product» the specification should contain information about authenticity of the cell line used in the BCP, namely: morphological characteristics, expression of specific markers, expression of specific genes, expression of specific proteins, as well as markers of cell line stability. At present Russia has no practical experience in BCP quality evaluation. The aim of the study was to substantiate methodological approaches to authentication of cell lines used in BCPs as illustrated by quality evaluation of the DF-2 model cell line using test methods that allow for characterisation of the morphological, genetic, immunophenotypic, and cytogenetic profile of the cell line. Materials and methods: the study analysed the DF-2 cell line — human dermal fibroblasts (mesenchymal stem cells) obtained from the Institute of Cytology of the Russian Academy of Sciences (St. Petersburg). The following analytical test methods were used in the study: morphological analysis; flow cytometry for immunophenotyping of the DF-2 model cell line; short tandem repeats for creating an allelic profile of the model cell line; cytogenetic analysis — differential DAPI staining of metaphase chromosomes. Results: the paper summarises methodological approaches to identification testing of medicines containing living human cells (BCP analogues) currently used in international practice, and presents the results of authentication of the model cell line. Conclusions: methods used for BCP identification testing should ensure unambiguous authentication of the cell line according to its specification. The study performed helped to work out the procedure of authentication of a model cell line.

Methodological aspects of the development of product files for biomedical cell products

“…Примером описания стратегии тестирования двухуровневой системы банков клеток является открытая документация на препараты (аналоги БМКП) Apligraf и Gintuit (Organogenisis) 37 , представляющие собой кератиноциты человека (верхний слой) и дермальные фибробласты (нижний слой) в матриксе из бычьего коллагена на пористой (поликарбонатной) мембране. Препараты Apligraf и Gintuit предназначены для регенерации кератинизированной ткани при синдроме диабетической стопы, варикозной язвы и устранения дефектов десен соответственно.…”

The Use of Short Tandem Repeat Analysis for Cell Line Authentication

“…В Российской Федерации на данный момент не существует наборов STR-анализа отечественного производства, которые были бы предназначены конкретно для определения подлинности КЛ человека. Ранее нами была показана возможность применения набора отечественного производителя COrDIS plus (ООО «Гордиз», Россия) для фрагментного анализа, содержащего одновременно 20 локусов из разных баз данных [18].…”