| RRC ID |
54902
|
| Author |
Kono K, Sawada R, Kuroda T, Yasuda S, Matsuyama S, Matsuyama A, Mizuguchi H, Sato Y.
|
| Title |
Development of selective cytotoxic viral vectors for concentration of undifferentiated cells in cardiomyocytes derived from human induced pluripotent stem cells.
|
| Journal |
Sci Rep
|
| Abstract |
Cell-processed therapeutic products (CTPs) derived from human pluripotent stem cells (hPSCs) have innovative applications in regenerative medicine. However, undifferentiated hPSCs possess tumorigenic potential; thus, sensitive methods for the detection of residual undifferentiated hPSCs are essential for the clinical use of hPSC-derived CTPs. The detection limit of the methods currently available is 1/105 (0.001%, undifferentiated hPSCs/differentiated cells) or more, which could be insufficient for the detection of residual hPSCs when CTPs contain more than 1 × 105 cells. In this study, we developed a novel approach to overcome this challenge, using adenovirus and adeno-associated virus (AdV and AAV)-based selective cytotoxic vectors. We constructed AdV and AAV vectors that possess a suicide gene, iCaspase 9 (iCasp9), regulated by the CMV promoter, which is dormant in hPSCs, for the selective expression of iCasp9 in differentiated cells. As expected, AdV/CMV-iCasp9 and AAV/CMV-iCasp9 exhibited cytotoxicity in cardiomyocytes but not in human induced pluripotent stem cells (hiPSCs). The vectors also induced apoptosis in hiPSC-derived cardiomyocytes, and the surviving cells exhibited higher levels of hPSC marker expression. These results indicate that the AdV- and AAV-based cytotoxic vectors concentrate cells expressing the undifferentiated cell markers in hiPSC-derived products and are promising biological tools for verifying the quality of CTPs.
|
| Volume |
9(1)
|
| Pages |
3630
|
| Published |
2019-3-6
|
| DOI |
10.1038/s41598-018-36848-5
|
| PII |
10.1038/s41598-018-36848-5
|
| PMID |
30842516
|
| PMC |
PMC6403330
|
| MeSH |
Adenoviridae / genetics*
Adenoviridae Infections / virology
Cell Differentiation*
Dependovirus / genetics*
Genetic Vectors / administration & dosage*
Genetic Vectors / genetics
Humans
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / metabolism
Induced Pluripotent Stem Cells / virology
Myocytes, Cardiac / metabolism
Myocytes, Cardiac / pathology*
Myocytes, Cardiac / virology
Parvoviridae Infections / virology
Regenerative Medicine*
|
| IF |
4.011
|
| Times Cited |
1
|
| Resource |
| Human and Animal Cells |
201B7(HPS0063) |
