Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNA
The reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) is gaining in importance in the fields of regenerative medicine, tissue engineering, and disease modeling. Patient-specific iPSCs have as an unlimited cell source a tremendous potential for generating various types of aut...
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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2019/7641767 |
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| author | Heidrun Steinle Marbod Weber Andreas Behring Ulrike Mau-Holzmann Christian Schlensak Hans Peter Wendel Meltem Avci-Adali |
| author_facet | Heidrun Steinle Marbod Weber Andreas Behring Ulrike Mau-Holzmann Christian Schlensak Hans Peter Wendel Meltem Avci-Adali |
| author_sort | Heidrun Steinle |
| collection | DOAJ |
| description | The reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) is gaining in importance in the fields of regenerative medicine, tissue engineering, and disease modeling. Patient-specific iPSCs have as an unlimited cell source a tremendous potential for generating various types of autologous cells. For the future clinical applicability of these iPSC-derived cells, the generation of iPSCs via nongenome integrating methods and the efficient reprogramming of patients’ somatic cells are required. In this study, 2 different RNA-based footprint-free methods for the generation of iPSCs were compared: the use of synthetic modified messenger RNAs (mRNAs) or self-replicating RNAs (srRNAs) encoding the reprogramming factors and GFP. Using both RNA-based methods, integration-free iPSCs without genomic alterations were obtained. The pluripotency characteristics identified by specific marker detection and the in vitro and in vivo trilineage differentiation capacity were comparable. Moreover, the incorporation of a GFP encoding sequence into the srRNA enabled a direct and convenient monitoring of the reprogramming procedure and the successful detection of srRNA translation in the transfected cells. Nevertheless, the use of a single srRNA to induce pluripotency was less time consuming, faster, and more efficient than the daily transfection of cells with synthetic mRNAs. Therefore, we believe that the srRNA-based approach might be more appropriate and efficient for the reprogramming of different types of somatic cells for clinical applications. |
| format | Article |
| id | doaj-art-3c28b5ebfe1a4b56bad6d4e55ac2596a |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-3c28b5ebfe1a4b56bad6d4e55ac2596a2025-02-03T07:23:49ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/76417677641767Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNAHeidrun Steinle0Marbod Weber1Andreas Behring2Ulrike Mau-Holzmann3Christian Schlensak4Hans Peter Wendel5Meltem Avci-Adali6University Hospital Tübingen, Department of Thoracic and Cardiovascular Surgery, Calwerstraße 7/1, 72076 Tübingen, GermanyUniversity Hospital Tübingen, Department of Thoracic and Cardiovascular Surgery, Calwerstraße 7/1, 72076 Tübingen, GermanyUniversity Hospital Tübingen, Department of Thoracic and Cardiovascular Surgery, Calwerstraße 7/1, 72076 Tübingen, GermanyUniversity Hospital Tübingen, Institute of Medical Genetics and Applied Genomics, Calwerstraße 7, 72076 Tübingen, GermanyUniversity Hospital Tübingen, Department of Thoracic and Cardiovascular Surgery, Calwerstraße 7/1, 72076 Tübingen, GermanyUniversity Hospital Tübingen, Department of Thoracic and Cardiovascular Surgery, Calwerstraße 7/1, 72076 Tübingen, GermanyUniversity Hospital Tübingen, Department of Thoracic and Cardiovascular Surgery, Calwerstraße 7/1, 72076 Tübingen, GermanyThe reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) is gaining in importance in the fields of regenerative medicine, tissue engineering, and disease modeling. Patient-specific iPSCs have as an unlimited cell source a tremendous potential for generating various types of autologous cells. For the future clinical applicability of these iPSC-derived cells, the generation of iPSCs via nongenome integrating methods and the efficient reprogramming of patients’ somatic cells are required. In this study, 2 different RNA-based footprint-free methods for the generation of iPSCs were compared: the use of synthetic modified messenger RNAs (mRNAs) or self-replicating RNAs (srRNAs) encoding the reprogramming factors and GFP. Using both RNA-based methods, integration-free iPSCs without genomic alterations were obtained. The pluripotency characteristics identified by specific marker detection and the in vitro and in vivo trilineage differentiation capacity were comparable. Moreover, the incorporation of a GFP encoding sequence into the srRNA enabled a direct and convenient monitoring of the reprogramming procedure and the successful detection of srRNA translation in the transfected cells. Nevertheless, the use of a single srRNA to induce pluripotency was less time consuming, faster, and more efficient than the daily transfection of cells with synthetic mRNAs. Therefore, we believe that the srRNA-based approach might be more appropriate and efficient for the reprogramming of different types of somatic cells for clinical applications.http://dx.doi.org/10.1155/2019/7641767 |
| spellingShingle | Heidrun Steinle Marbod Weber Andreas Behring Ulrike Mau-Holzmann Christian Schlensak Hans Peter Wendel Meltem Avci-Adali Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNA Stem Cells International |
| title | Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNA |
| title_full | Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNA |
| title_fullStr | Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNA |
| title_full_unstemmed | Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNA |
| title_short | Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNA |
| title_sort | generation of ipscs by nonintegrative rna based reprogramming techniques benefits of self replicating rna versus synthetic mrna |
| url | http://dx.doi.org/10.1155/2019/7641767 |
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