Reference - Detail
| RRC ID | 71728 |
|---|---|
| Author | Yasui R, Matsui A, Sekine K, Okamoto S, Taniguchi H. |
| Title | Highly Sensitive Detection of Human Pluripotent Stem Cells by Loop-Mediated Isothermal Amplification. |
| Journal | Stem Cell Rev Rep |
| Abstract |
For safe regenerative medicines, contaminated or remaining tumorigenic undifferentiated cells in cell-derived products must be rigorously assessed through sensitive assays. Although in vitro nucleic acid tests offer particularly sensitive tumorigenicity-associated assays, the human pluripotent stem cell (hPSC) detectability is partly constrained by the small input amount of RNA per test. To overcome this limitation, we developed reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays that are highly gene specific and robust against interfering materials. LAMP could readily assay microgram order of input sample per test and detected an equivalent model of 0.00002% hiPSC contamination in a simple one-pot reaction. For the evaluation of cell-derived total RNA, RT-LAMP detected spiked-in hPSCs among hPSC-derived trilineage cells utilizing multiple pluripotency RNAs. We also developed multiplex RT-LAMP assays and further applied for in situ cell imaging, achieving specific co-staining of pluripotency proteins and RNAs. Our attempts uncovered the utility of RT-LAMP approaches for tumorigenicity-associated assays, supporting practical applications of regenerative medicine. |
| Volume | 18(8) |
| Pages | 2995-3007 |
| Published | 2022-12-1 |
| DOI | 10.1007/s12015-022-10402-3 |
| PII | 10.1007/s12015-022-10402-3 |
| PMID | 35661077 |
| PMC | PMC9622575 |
| MeSH | Humans Nucleic Acid Amplification Techniques* / methods Pluripotent Stem Cells* RNA Sensitivity and Specificity |
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 3 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | HiPS-RIKEN-2A(HPS0009) HiPS-RIKEN-12A(HPS0029) Nips-B2(HPS0223) 293T(RCB2202) |