Generation of a human induced pluripotent stem cell line, KSCBi003-A, from human adipose tissue-derived mesenchymal stem cells using a chromosomal integration-free system

Choi, Hye Young; Kim, Sojung; Go, Gue Youn; Kwon, Ara; Im, Young Sam; Ha, Hye-Yeong; Hong, Jin Tae; Jung, Ji‐Won; Koo, Soo Kyung

doi:10.1016/j.scr.2018.06.003

Cited by 2 publications

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“…It seems that major determinants of reprogramming success are sample-related factors, cell of origin ( proliferation and differentiation status of cell and cellular epigenetic features ), and culture conditions rather than the method itself [28]. A range of somatic cell types including neonatal fibroblasts, dental pulp cells, adipose cells, CD34+ cells from the umbilical cord, and peripheral blood mononuclear cells has been successfully reprogrammed with SeV and Episomal methods [11, 17, 29, 30]. We preferred to reprogram BM-MSC of osteopetrosis patients, because of their multipotency, ease of purification/expansion and well-established characteristics [19, 20].…”

Section: Discussionmentioning

confidence: 99%

Osteopetrotic induced pluripotent stem cells derived from patients with different disease-associated mutations by non-integrating reprogramming methods

et al. 2019

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Background Autosomal recessive osteopetrosis is a genetically and phenotypically heterogeneous disease, caused by defects in osteoclast formation and function. The only available treatment is allogeneic stem cell transplantation that has still high morbidity and mortality. The goal of the present study was to generate iPSCs from bone marrow-derived MSCs of osteopetrosis patients with three most common mutations by using two different integration-free gene transfer methods and compare their efficiencies. The secondary objective was to select the most appropriate integration-free production method for our institutional iPSC bank using this rare disease as a prototype. Methods Two different integration-free gene transfer methods (episomal and Sendai viral vectors) were tested and compared on the same set of patient samples exhibiting three different mutations associated with osteopetrosis. Generated iPSCs were characterized by standard assays, including immunophenotyping, immunocytochemistry, RT-PCR, embryoid body, and teratoma assays. Karyotype analyses were performed to evaluate genetic stability. Results iPSC lines exhibiting typical ESC-like colony morphology were shown to express pluripotency markers by immunofluorescence staining. Over 90% of the cells were found positive for SSEA-4 and OCT3/4 and negative/weak positive for CD29 by flow cytometry. Immunohistochemical staining of teratoma and spontaneously differentiated embryoid body sections confirmed their trilineage differentiation potential. All iPSC lines expressed pluripotency-related genes. Karyotype analyses were found normal. Direct sequencing of PCR-amplified DNA showed that disease-related mutations were retained in the patient-specific iPSCs. Conclusion Generation of iPSC using SeV and episomal DNA vectors have several advantages over other methods like the ease of production, reliability, high efficiency, and safety, which is required for translational research. Furthermore, owing to the pluripotency and self-renewal capacity, patient-specific iPSCs seem to be ideal cell source for the modeling of a rare genetic bone disease like osteopetrosis to identify osteoclast defects, leading to clinical heterogeneity in osteopetrosis patients, especially among those with different mutations in the same gene. Electronic supplementary material The online version of this article (10.1186/s13287-019-1316-8) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Osteopetrotic induced pluripotent stem cells derived from patients with different disease-associated mutations by non-integrating reprogramming methods

et al. 2019

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Human iPSC banking: barriers and opportunities

et al. 2019

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The introduction of induced pluripotent stem cells (iPSCs) has opened up the potential for personalized cell therapies and ushered in new opportunities for regenerative medicine, disease modeling, iPSC-based drug discovery and toxicity assessment. Over the past 10 years, several initiatives have been established that aim to collect and generate a large amount of human iPSCs for scientific research purposes. In this review, we compare the construction and operation strategy of some iPSC banks as well as their ongoing development. We also introduce the technical challenges and offer future perspectives pertaining to the establishment and management of iPSC banks.

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Generation of a human induced pluripotent stem cell line, KSCBi003-A, from human adipose tissue-derived mesenchymal stem cells using a chromosomal integration-free system

Cited by 2 publications

References 4 publications

Osteopetrotic induced pluripotent stem cells derived from patients with different disease-associated mutations by non-integrating reprogramming methods

Osteopetrotic induced pluripotent stem cells derived from patients with different disease-associated mutations by non-integrating reprogramming methods

Human iPSC banking: barriers and opportunities

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