Development, characterization, and hematopoietic differentiation of Griscelli syndrome type 2 induced pluripotent stem cells
| Autor: | Gülben Gürhan Sevinç, Elif Bilgiç, Özgür Doğuş Erol, Fatima Aerts-Kaya, Aysen Gunel-Ozcan, Tamer T. Onder, Duygu Uckan-Cetinkaya, Burcu Pervin, Petek Korkusuz, Gülen Güney-Esken |
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| Přispěvatelé: | Sevinç, Gülben Gürhan, Önder, Tamer Tevfik (ORCID 0000-0002-2372-9158 & YÖK ID 42946), Güney-Esken, Gülen, Erol, Özgür Doğuş, Pervin, Burcu, Korkusuz, Petek, Günel-Özcan, Ayşen, Uçkan-Çetinkaya, Duygu, Aerts-Kaya, Fatima, Koç University Research Center for Translational Medicine (KUTTAM) / Koç Üniversitesi Translasyonel Tıp Araştırma Merkezi (KUTTAM), Graduate School of Health Sciences, School of Medicine |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Homeobox protein NANOG
Medicine (General) Primary Immunodeficiency Diseases Mesenchymal stromal cells Medicine (miscellaneous) QD415-436 Gene mutation Biology Biochemistry Genetics and Molecular Biology (miscellaneous) Biochemistry Lymphohistiocytosis Hemophagocytic Kruppel-Like Factor 4 R5-920 SOX2 Humans Bone marrow Progenitor cell Induced pluripotent stem cell Griscelli syndrome type 2 Hematopoietic stem cells Induced pluripotent stem cells Research Mesenchymal stem cell Hematopoietic Stem Cell Transplantation Cell Biology Research and experimental medicine Feeder Cells Cell Differentiation Piebaldism Cell biology Haematopoiesis Molecular Medicine Stem cell |
| Zdroj: | Stem Cell Research & Therapy, Vol 12, Iss 1, Pp 1-13 (2021) Stem Cell Research & Therapy |
| ISSN: | 1757-6512 |
| Popis: | Background: Griscelli syndrome type 2 (GS-2) is a rare, autosomal recessive immune deficiency syndrome caused by a mutation in the RAB27A gene, which results in the absence of a protein involved in vesicle trafficking and consequent loss of function of in particular cytotoxic T and NK cells. Induced pluripotent stem cells (iPSC) express genes associated with pluripotency, have the capacity for infinite expansion, and can differentiate into cells from all three germ layers. They can be induced using integrative or non-integrative systems for transfer of the Oct4, Sox2, Klf4, and cMyc (OSKM) transcription factors. To better understand the pathophysiology of GS-2 and to test novel treatment options, there is a need for an in vitro model of GS-2. Methods: here, we generated iPSCs from 3 different GS-2 patients using lentiviral vectors. The iPSCs were characterized using flow cytometry and RT-PCR and tested for the expression of pluripotency markers. In vivo differentiation to cells from all three germlines was tested using a teratoma assay. In vitro differentiation of GS-2 iPSCs into hematopoietic stem and progenitor cells was done using Op9 feeder layers and specified media. Results: all GS-2 iPSC clones displayed a normal karyotype (46XX or 46XY) and were shown to express the same RAB27A gene mutation that was present in the original somatic donor cells. GS-2 iPSCs expressed SSEA1, SSEA4, TRA-1-60, TRA-1-81, and OCT4 proteins, and SOX2, NANOG, and OCT4 expression were confirmed by RT-PCR. Differentiation capacity into cells from all three germ layers was confirmed using the teratoma assay. GS-2 iPSCs showed the capacity to differentiate into cells of the hematopoietic lineage. Conclusions: using the lentiviral transfer of OSKM, we were able to generate different iPSC clones from 3 GS-2 patients. These cells can be used in future studies for the development of novel treatment options and to study the pathophysiology of GS-2 disease. Scientific and Technological Research Council of Turkey (TÜBİTAK) |
| Databáze: | OpenAIRE |
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