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| Status |
Public on Jan 06, 2021 |
| Title |
Robust parameter design of human induced pluripotent stem cell differentiation protocols defines lineage‐specific induction of anterior‐posterior gut tube endodermal cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Tissues and cells derived from pluripotent stem cells (PSC) are likely to become widely used in disease modeling, drug screening, and regenerative medicine. For these applications, the in vitro PSC differentiation process must be elaborately investigated and controlled to reliably obtain the desired end products. However, because traditional experimental methods, such as one factor at a time (OFAT) or brute-force approaches, are impractical for detailed screening of complex PSC cultivation conditions, more strategic and effective screening based on statistical design of experiments (DOE) ought to be indispensable. Among various DOE approaches, we regard robust parameter design (RPD) as particularly suited for differentiation protocol optimization due to its suitability for multi-factorial screening. We confirmed the adaptability of RPD for investigating human induced PSC (hiPSC) lineage specification toward anterior–posterior gut tube endodermal cells and clarified the contribution of each cell signaling pathway and the effect of cell signaling condition alteration on marker RNA expression levels, while increasing the efficiency of the screening in 243-fold (18 vs 4374) compared to that of a brute-force approach. Specific induction of anterior foregut, hepatic, pancreatic, or mid-hindgut cells was achieved using seven iPSC strains with the optimal culture protocols established on the basis of RPD analysis. RPD has the potential to enable efficient construction and optimization of PSC differentiation protocols, and its use is recommended from fundamental research to mass production of PSC-derived products.
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| Overall design |
mRNA expression profiles of iPSC-derived anterior foregut, pancreatic, hepatic, and mid-hindgut cells
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| Contributor(s) |
Sekine K, Yasui R, Yamaguchi K |
| Citation(s) |
33400835 |
| Submission date |
Dec 14, 2020 |
| Last update date |
Jan 06, 2021 |
| Contact name |
Keisuke Sekine |
| Organization name |
National Cancer Center Research Institute, Japan
|
| Department |
Graduate School of Medicine
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| Lab |
Laboratory of Cancer Cell Systems
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| Street address |
5-1-1 Tsukiji
|
| City |
Chuo-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
104-0045 |
| Country |
Japan |
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| Platforms (1) |
| GPL17303 |
Ion Torrent Proton (Homo sapiens) |
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| Samples (28)
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| Relations |
| BioProject |
PRJNA685108 |
| SRA |
SRP297873 |
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