Abstract
Differentiation of induced pluripotent stem cells (iPSCs) from patients and healthy donors allows in vitro study of genetic disorders. The authors have previously reported a clinical case of recurrent pregnancy loss in a patient with skewed X-chromosome inactivation in peripheral blood lymphocytes, endometrium, and buccal epithelium. A 239 kb microdeletion at Xq24 that affected eight genes, including UBE2A, has been found. In this work, an iPSC line iTAF15Xsk4 was produced from the patient’s skin fibroblasts using nonintegrating episomal vectors. The iPSC line had a normal karyotype, expressed pluripotency markers, and expressed markers of all three germ layers upon differentiation in embryoid bodies. This cell line could be used for the UBE2A deficiency syndrome study.
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ACKNOWLEDGMENTS
The generation and cultivation of iPSCs were carried out at the Collective Center of the Institute of Cytology and Genetics (Siberian Branch, Russian Academy of Sciences) “Collection of Pluripotent Human and Mammalian Cell Cultures for Biological and Biomedical Research” (https://ckp.icgen.ru/cells/; http://www.biores.cytogen.ru/brc_cells/collections/ICG_SB_RAS_CELL). Visualization and microscopic analysis of the immunofluorescence staining results were carried out at the Public Center for Microscopy (Siberian Branch, Russian Academy of Sciences) (https://ckp.icgen.ru/ckpmabo/), supported by the budget project of the Institute of Cytology and Genetics (Siberian Branch, Russian Academy of Sciences) FWNR-2022-0019. Obtaining a skin biopsy and cultivation of primary fibroblasts was carried out at the Medical Genomic Collective Center of the Research Institute of Medical Genetics, Tomsk.
Funding
The work was carried out with the financial support of the Ministry of Science and Higher Education of the Russian Federation (agreement no. 075-15-2021-1063 dated September 28, 2021). Primary skin fibroblasts were obtained with the financial support of the Russian Science Foundation, grant no. 21-65-00017.
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A.G. Menzorov reprogrammed fibroblasts and generated the iPSC line. I.E. Pristyazhnyuk performed cytogenetic analysis. N.I. Meshcheryakov performed DNA and RNA isolation, PCR and RT-PCR analysis. T.V. Nikitina cultivated primary fibroblasts. A.A. Kashevarova and D.A. Fedotov performed RT-PCR. E.N. Tolmacheva, L.I. Minaycheva, and L.P. Nazarenko provided medical support to the patient and performed a skin biopsy. A.G. Menzorov and I.N. Lebedev designed the experiment, analyzed the obtained data, and participated in writing the article. All the authors participated in the discussion of the results.
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The authors of this work declare that they have no conflicts of interest.
ETHICS APPROVAL AND CONSENT TO PARTICIPATE
The study was approved by the Ethical Commission of the Research Institute of Medical Genetics (Tomsk National Research Medical Center, Russian Academy of Sciences) (protocol no. 10 of February 15, 2021). The patient signed a voluntary informed consent with her own hand.
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Passport of the Pluripotent Stem Cell Lines
Translated by E. Tolkunova
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Pristyazhnyuk, I.E., Meshcheryakov, N.I., Nikitina, T.V. et al. Generation of Induced Pluripotent Stem Cell Line iTAF15Xsk4 from Fibroblasts of a Patient with Microdeletion at Xq24. Russ J Dev Biol 54, 358–364 (2023). https://doi.org/10.1134/S1062360423060073
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DOI: https://doi.org/10.1134/S1062360423060073