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A physical biomarker of the quality of cultured corneal endothelial cells and of the long-term prognosis of corneal restoration in patients


Dysfunction of the corneal endothelium reduces the transparency of the cornea and can cause blindness. Because corneal endothelial cells have an extremely limited proliferative ability in vivo, treatment for corneal endothelial dysfunction involves the transplantation of donor corneal tissue. Corneal endothelium can also be restored via intraocular injection of endothelial cells in suspension after their expansion in vitro. Yet, because quality assessment during the expansion of the cells is a destructive process, a substantial number of the cultured cells are lost. Here, we show that the ‘spring constant’ of the effective interaction potential between endothelial cells in a confluent monolayer serves as a biomarker of the quality of corneal endothelial cells in vitro and of the long-term prognosis of corneal restoration in patients treated with culture-expanded endothelial cells or with transplanted corneas. The biomarker can be measured from phase contrast imaging in vitro and from specular microscopy in vivo, and may enable a shift from passive monitoring to pre-emptive intervention in patients with severe corneal disorders.

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Fig. 1: Restoration of the human cornea via injection of culture-expanded corneal endothelial cells.
Fig. 2: Course of experiments and analyses in vitro and in vivo.
Fig. 3: Characterization of in vivo corneal endothelium.
Fig. 4: Quality control of in vitro cell sources.
Fig. 5: Classification accuracies of different indicators.
Fig. 6: Collective order for predictive diagnosis.

Data availability

The authors declare that the main data supporting the results in this study are available within the paper and its Supplementary Information. The raw and analysed datasets generated during the study are available for research purposes from the corresponding authors on reasonable request.

Code availability

All Igor codes in this work are available from the corresponding authors on reasonable request.


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We thank the Baptist Eye Institute for access to the clinical records, H. Nakagawa for data collection, K. Yoshikawa for stimulating discussions, and J. Bush for reviewing the manuscript. This work was supported by Nakatani Foundation (M.Tanaka and M.U.), JSPS (17H00855 to M.Tanaka; 16K05515 to A.Y.), MEXT (16KT0070 to M.Tanaka), the Highway Program for Realization of Regenerative Medicine of the Japan Agency for Medical Research and Development (S.K.) and the Research Project for Practical Applications of Regenerative Medicine of the Japanese Ministry of Health, Labor and Welfare (S.K.).

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Authors and Affiliations



M.U. and M.Tanaka conceived and directed the research. A.Y. and H.T. performed the research and analysis. M.Toda collected the data. C.S. and S.K. supervised the retrospective analysis of clinical research. A.Y., H.T., J.H., S.K., M.U. and M.Tanaka wrote the paper.

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Correspondence to Morio Ueno or Motomu Tanaka.

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The authors declare no competing interests.

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Yamamoto, A., Tanaka, H., Toda, M. et al. A physical biomarker of the quality of cultured corneal endothelial cells and of the long-term prognosis of corneal restoration in patients. Nat Biomed Eng 3, 953–960 (2019).

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