Development of Cell Analysis Software for Cultivated Corneal Endothelial Cells

Okumura, Naoki; Ishida, Naoya; Kakutani, Kazuya; Hongo, Akane; Hiwa, Satoru; Hiroyasu, Tomoyuki; Koizumi, Noriko

doi:10.1097/ico.0000000000001317

Cited by 7 publications

(1 citation statement)

References 26 publications

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“…This approval will allow HCECs to be marketed as a product, thereby eventually allowing physicians and patients worldwide to access this new therapy. We are currently optimizing the entire protocol, from improving the efficiency of in vivo expansion to establishment of large-scale commercial cell culture protocols, transportation methods, quality control practices, and cryopreservation procedures to enable the CPC to manufacture and provide HCECs as a product [16–18].…”

Section: Introductionmentioning

confidence: 99%

Feasibility of a cryopreservation of cultured human corneal endothelial cells

et al. 2019

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Transparency of the cornea is essential for vision and is maintained by the corneal endothelium. Consequently, corneal endothelial decompensation arising from irreversible damage to the corneal endothelium causes severe vision impairment. Until recently, transplantation of donor corneas was the only therapeutic choice for treatment of endothelial decompensation. In 2013, we initiated clinical research into cell-based therapy involving injection of a suspension of cultured human corneal endothelial cells (HCECs), in combination with Rho kinase inhibitor, into the anterior chamber. The aim of the present study was to establish a protocol for cryopreservation of HCECs to allow large-scale commercial manufacturing of these cells. This study focused on the effects of various cryopreservation reagents on HCEC viability. Screening of several commercially available cryopreservation reagents identified Bambanker hRM as an effective agent that maintained a cell viability of 89.4% after 14 days of cryopreservation, equivalent to the cell viability of 89.2% for non-cryopreserved control cells. The use of Bambanker hRM and HCECs at a similar grade to that used clinically for cell based therapy (passage 3–5 and a cell density higher than 2000 cells/mm 2 ) gave a similar cell density for cryopreserved HCECs to that of non-preserved control HCECs after 28 days of cultivation (2099 cells/mm 2 and 2111 cells/mm 2 , respectively). HCECs preserved using Bambanker hRM grew in a similar fashion to non-preserved control HCECs and formed a monolayer sheet-like structure. Cryopreservation of HCECs has multiple advantages including the ability to accumulate stocks of master cells, to transport HCEC stocks, and to manufacture HCECs on demand for use in cell-based treatment of endothelial decompensation.

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Section: Introductionmentioning

confidence: 99%