2020 Fiscal Year Research-status Report
The establishment of a biomimetic human iPSC-derived cornea-on-a-chip for the pre-clinical evaluation of ophthalmic nanomedicines
| Project/Area Number |
20K20168
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Project Period (FY) |
2020-04-01 – 2022-03-31
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| Keywords | Cornea / hPSCs / Microfluidic / Nano toxicity |
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| Outline of Annual Research Achievements |
The applicant has developed a protocol for the generation of human corneal epithelial cells from human pluripotent stem cells (hPSCs) using a serum-free and xeno-free culture system supplemented with small chemicals. The applicant has succeeded in the generation of hPSC-derived corneal epithelial cells characterized with significant expression of the eye ectoderm developmental marker and the corneal epithelial maturation marker in comparison with a conventional human corneal epithelial cell line (HCE-T). Moreover, the applicant has optimized the necessary culturing conditions of hPSC-derived corneal epithelial cells before their seeding in a microfluidic device. Meanwhile, the applicant has improved the flow system for the generation of blinking-like stimulus using an air pumping system.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
In general, the research is progressing according to the initial implementation plan. The protocol for corneal epithelial cells generation from hPSCs is completed and the optimization of cell culture conditions such as cells density and type of extracellular matrix (ECM) needed for the formation of the corneal epithelium in the microfluidic device are performed.
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| Strategy for Future Research Activity |
1-Determine the function of the hPSC-derived corneal epithelium barrier on a chip using gene expression analysis.
2-Leverage the microfluidic device based on the applicant previous publication to be used for the evaluation of molecules secretion and transportation across the corneal epithelial barrier to determine the spatiotemporal nanotoxicity.
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| Causes of Carryover |
As the applicant has changed the research institution, consumables for the fabrication of microfluidic devices, culture media and reagents for the cultivation and differentiation of human pluripotent stem cells are to be included. Besides, due to the COVID-19 pandemic, the applicant could not use the budget for attending international or domestic meetings effectively. Moreover, as this is the last year of the project, the applicant plans to report the results in the form of a research paper based on the experimental results , so the costs associated with this are included. Therefore, the applicant is expecting to use the budget in the current fiscal year for the above-mentioned purposes.
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Research Products
(5 results)
