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Vol 56(2022) N 2 p. 276-282; DOI 10.1134/S0026893322020030 Full Text

A.A. Babovskaya1*, E.A. Trifonova1,2, V.N. Serebrova1, M.G. Svarovskaya1,2, A.A. Zarubin1, O.V. Zhilyakova2, T.V. Gabidulina2, A.A. Poltanova2, L.V. Rychkova3, V.A. Stepanov1

Protocol of Transcriptome Analysis of Decidual Placenta Cells

1Research Institute of Medical Genetics, Tomsk National Research Medical Center, Tomsk, 634050 Russia
2Siberian State Medical University, Tomsk, 634050 Russia
3Scientific Center for Family Health and Human Reproduction Problems, Irkutsk, 664003 Russia

*anastasia.babovskaya@medgenetics.ru
Received - 2021-07-02; Revised - 2021-09-29; Accepted - 2021-10-13

The advent of high-throughput sequencing technologies has expanded our understanding of the biological significance of non-coding regions of the genome. In recent years, more and more studies have been devoted to studying the role of noncoding RNAs in the development of diseases, as well as their participation in various cellular processes. Until now, all transcriptome studies of native placental tissue with the description of the noncoding RNA region were carried out without isolating individual cell populations. This approach, due to the high cellular heterogeneity of the placental tissue, significantly complicates the ability to determine the molecular-biological functions of individual cells and their role in the molecular pathogenesis of reproductive disorders. In this work, we propose a technique for obtaining total RNA from single decidual cells of frozen placental tissue obtained by laser-capture microdissection technology for transcriptome sequencing, including a cluster of noncoding RNAs. This technique can be successfully used to study the full-genome expression profile of other placental cell populations. The high accuracy of results on the transcriptome profiling of decidual cells obtained using the developed technique was additionally confirmed by an integrative analysis with the results of a 10x Genomics experiment.

placenta, decidual cells, laser-capture microdissection, transcriptomics, RNA, high-throughput sequencing technologies



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