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| Status |
Public on May 21, 2019 |
| Title |
Interrupted reprogramming into induced pluripotent stem cells does not rejuvenate human mesenchymal stromal cells |
| Organism |
Homo sapiens |
| Experiment type |
SNP genotyping by SNP array
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| Summary |
Replicative senescence hampers application of mesenchymal stromal cells (MSCs) because it limits culture expansion, impairs differentiation potential, and hinders reliable standardization of cell products. MSCs can be rejuvenated by reprogramming into induced pluripotent stem cells (iPSCs), which is associated with complete erasure of age- and senescence-associated DNA methylation (DNAm) patterns. However, this process is also associated with erasure of cell-type and tissue-specific epigenetic characteristics that are not recapitulated upon re-differentiation towards MSCs. In this study, we therefore followed the hypothesis that overexpression of pluripotency factors under culture conditions that do not allow full reprogramming might reset senescence-associated changes without entering a pluripotent state. MSCs were transfected with episomal plasmids and either successfully reprogrammed into iPSCs or cultured in different media with continuous passaging every week. Overexpression of pluripotency factors without reprogramming did neither prolong culture expansion nor ameliorate molecular and epigenetic hallmarks of senescence. Notably, transfection resulted in immortalization of one cell preparation with gain of large parts of the long arm of chromosome 1. Taken together, premature termination of reprogramming does not result in rejuvenation of MSCs and harbours the risk of transformation. This approach is therefore not suitable to rejuvenate cells for cellular therapy.
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| Overall design |
The Affymetrix CytoScan® HD Array was applied on DNA extracted from transfected human MSCs of donor 2 at passage 4 and 12 because cells displayed altered morphology and immunophenotype at late passage. Consequently, we performed copy number variation (CNV) analysis to look for possible contaminations with other cell lines or chromosomal aberrations at late passage.
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| Contributor(s) |
Goebel C, Goetzke R, Eggermann T, Wagner W |
| Citation(s) |
30076334 |
| Submission date |
Jun 12, 2018 |
| Last update date |
May 21, 2019 |
| Contact name |
Wolfgang Wagner |
| E-mail(s) |
wwagner@ukaachen.de
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| Phone |
+49 241 8088611
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| Organization name |
RWTH Aachen University
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| Department |
Helmholtz Institute for Biomedical Engineering
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| Lab |
Stem Cell Biology and Cellular Engineering
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| Street address |
Pauwelsstrasse 20
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| City |
Aachen |
| ZIP/Postal code |
52074 |
| Country |
Germany |
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| Platforms (1) |
| GPL16131 |
[CytoScanHD_Array] Affymetrix CytoScan HD Array |
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| Samples (2) |
| GSM3186894 |
BM-MSCs_donor 2_transfected_hPL-medium_P4 |
| GSM3186895 |
BM-MSCs_donor 2_transfected_hPL-medium_P12 |
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| Relations |
| BioProject |
PRJNA475735 |
