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A High-Throughput Method as a Diagnostic Tool for HIV Detection in Patient-Specific Induced Pluripotent Stem Cells Generated by Different Reprogramming Methods
Joint Authors
Dressel, Ralf
Hübscher, Daniela
Rebs, Sabine
Haupt, Luis
Borchert, Thomas
Guessoum, Celina Isabell
Treu, Franziska
Köhne, Steffen
Maus, Andreas
Hambrecht, Mario
Sossalla, Samuel
Uy, Angela
Jakob, Mark
Hasenfuss, Gerd
Streckfuss-Bömeke, Katrin
Source
Issue
Vol. 2019, Issue 2019 (31 Dec. 2019), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2019-08-05
Country of Publication
Egypt
No. of Pages
11
Abstract EN
Induced pluripotent stem cells (iPSCs) provide a unique opportunity for generation of patient-specific cells for use in translational purposes.
We aimed to compare iPSCs generated by different reprogramming methods regarding their reprogramming efficiency, pluripotency capacity, and the possibility to use high-throughput PCR-based methods for detection of human pathogenic viruses.
iPSCs from skin fibroblasts (FB), peripheral blood mononuclear cells (PBMCs), or mesenchymal stem cells (MSCs) were generated by using three different reprogramming systems including chromosomal integrating and nonintegrating methods.
Reprogramming efficiencies were in accordance with the literature, indicating that the parental cell type and the reprogramming method play a major role for the reprogramming efficiencies (FB: STEMCCA: 1.30±0.18, Sendai virus: 1.37±0.01, and episomal plasmids: 0.04±0.02; PBMCs: Sendai virus: 0.002±0.001, episomal plasmids: 0) but result in the same characteristics of pluripotency.
We found the highest reprogramming efficiencies for MSC with 3.32±1.2 by using episomal plasmids.
Since GMP standard working procedures and screening units need virus contamination-free cell lines, we studied HIV-1 contamination in the generated iPSCs.
We used the high-throughput cobas® 6800/8800 system, which is normally used for detection of HIV-1 in plasma of patients, and found that footprint-free reprogramming methods as episomal plasmids and Sendai virus are useful for the described virus detection method.
This fast, cost-effective, robust, and reliable assay demonstrates the feasibility to use high-throughput PCR-based methods for detection of human pathogenic viruses in ps-iPSC lines that were generated with nongenome integrating reprogramming methods.
American Psychological Association (APA)
Hübscher, Daniela& Rebs, Sabine& Haupt, Luis& Borchert, Thomas& Guessoum, Celina Isabell& Treu, Franziska…[et al.]. 2019. A High-Throughput Method as a Diagnostic Tool for HIV Detection in Patient-Specific Induced Pluripotent Stem Cells Generated by Different Reprogramming Methods. Stem Cells International،Vol. 2019, no. 2019, pp.1-11.
https://search.emarefa.net/detail/BIM-1208401
Modern Language Association (MLA)
Hübscher, Daniela…[et al.]. A High-Throughput Method as a Diagnostic Tool for HIV Detection in Patient-Specific Induced Pluripotent Stem Cells Generated by Different Reprogramming Methods. Stem Cells International No. 2019 (2019), pp.1-11.
https://search.emarefa.net/detail/BIM-1208401
American Medical Association (AMA)
Hübscher, Daniela& Rebs, Sabine& Haupt, Luis& Borchert, Thomas& Guessoum, Celina Isabell& Treu, Franziska…[et al.]. A High-Throughput Method as a Diagnostic Tool for HIV Detection in Patient-Specific Induced Pluripotent Stem Cells Generated by Different Reprogramming Methods. Stem Cells International. 2019. Vol. 2019, no. 2019, pp.1-11.
https://search.emarefa.net/detail/BIM-1208401
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1208401