Int J Stem Cells.
2023 Feb;16(1):36-43. 10.15283/ijsc22177.
Generation of Induced Pluripotent Stem Cells from Lymphoblastoid Cell Lines by Electroporation of Episomal Vectors
- Affiliations
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- 1Department of Medical Life Sciences, College of Medicine, The Catholic University of Korea, Seoul, Korea
- 2School of Biotechnology and Healthcare, Wuyi University, Jiangmen, China
- 3Department of General Pediatrics, University of Children’s Hospital Muenster, Muenster, Germany
- 4Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany
- 5Department of Cardiac Development and Remodelling, Max-Planck-Institute for Heart and Lung Research, Bad Nauheim, Germany
- KMID: 2539619
- DOI: http://doi.org/10.15283/ijsc22177
Abstract
- Background and Objectives
Lymphoblastoid cell lines (LCLs) deposited from disease-affected individuals could be a valuable donor cell source for generating disease-specific induced pluripotent stem cells (iPSCs). However, generation of iPSCs from the LCLs is still challenging, as yet no effective gene delivery strategy has been developed.
Methods and Results
Here, we reveal an effective gene delivery method specifically for LCLs. We found that LCLs appear to be refractory toward retroviral and lentiviral transduction. Consequently, lentiviral and retroviral transduction of OCT4, SOX2, KFL4 and c-MYC into LCLs does not elicit iPSC colony formation. Interestingly, however we found that transfection of oriP/EBNA-1-based episomal vectors by electroporation is an efficient gene delivery system into LCLs, enabling iPSC generation from LCLs. These iPSCs expressed pluripotency makers (OCT4, NANOG, SSEA4, SALL4) and could form embryoid bodies.
Conclusions
Our data show that electroporation is an effective gene delivery method with which LCLs can be efficiently reprogrammed into iPSCs.
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