Self-organization, quality control, and preclinical studies of human iPSC-derived retinal sheets for tissue-transplantation therapy

Watari, K.; Yamasaki, S.; Tu, H.-Y.; Shikamura, M.; Kamei, T.; Adachi, H.; Tochitani, T.; Kita, Y.; Nakamura, A.; Ueyama, K.; Ono, K.; Morinaga, C.; Matsuyama, T.; Sho, J.; Nakamura, M.; Fujiwara, M.; Hori, Y.; Tanabe, A.; Hirai, R.; Terai, O.; Ohno, O.; Ohara, H.; Hayama, T.; Ikeda, A.; Nukaya, D.; Matsushita, K.; Takahashi, M.; Kishino, A.; Kimura, T.; Kawamata, S.; Mandai, M.; Kuwahara, A.

doi:10.1101/2022.02.18.480968

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bioRxiv · 10.1101/2022.02.18.480968

Self-organization, quality control, and preclinical studies of human iPSC-derived retinal sheets for tissue-transplantation therapy

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Abstract

Three-dimensional retinal organoids (3D-retinas) are a promising graft source for transplantation therapy. We previously developed self-organizing culture for 3D-retina generation from human pluripotent stem cells (hPSCs). Here we present a quality control method and preclinical studies for tissue-sheet transplantation. Self-organizing hPSCs differentiated into both retinal and off-target tissues. Gene expression analyses identified the major off-target tissues as eye-related, cortex-like, and spinal cord-like tissues. For quality control, we developed a qPCR-based test in which each hPSC-derived neuroepithelium was dissected into two tissue-sheets: inner-central sheet for transplantation and outer-peripheral sheet for qPCR to ensure retinal tissue selection. During qPCR, tissue-sheets were stored for 3-4 days using a newly developed preservation method. In a rat tumorigenicity study, no transplant-related adverse events were observed. In retinal degeneration model rats, retinal transplants differentiated into mature photoreceptors and exhibited light responses in electrophysiology assays. These results demonstrate our rationale toward self-organizing retinal sheet transplantation therapy.

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Watari, K., Yamasaki, S., Tu, H.-Y., Shikamura, M., Kamei, T., Adachi, H., Tochitani, T., Kita, Y., Nakamura, A., Ueyama, K., Ono, K., Morinaga, C., Matsuyama, T., Sho, J., Nakamura, M., Fujiwara, M., Hori, Y., Tanabe, A., Hirai, R., Terai, O., Ohno, O., Ohara, H., Hayama, T., Ikeda, A., Nukaya, D., Matsushita, K., Takahashi, M., Kishino, A., Kimura, T., Kawamata, S., Mandai, M., Kuwahara, A.. 2022-02-19. Self-organization, quality control, and preclinical studies of human iPSC-derived retinal sheets for tissue-transplantation therapy. https://doi.org/10.1101/2022.02.18.480968

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