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Feasibility of a cryopreservation of cultured human corneal endothelial cells

Naoki Okumura, Takato Kagami, Kyoko Watanabe, Saori Kadoya, Masakazu Sato, Noriko KoizumiID* Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan 

The cornea plays an essential role in human vision. Turbidity of the cornea can cause a reduction of its function. this reduction is normally caused by severe damage to the colonial endothelium. The group of Koizumi et al. has started a clinical trial using human corneal endothelial cells to replace the cells. These cells could be a therapeutic solution to regain the corneal transparency. The clinical trial depends, how ever, on cryopreserved cells before adding the cells to the patient. In this paper they compared the efficiency in cryopreserving cells using various cryopreservation media. Their conclusion is that Bambanker hRM is the best cryopreservation medium!

Here are some of the reasons why they chose Bambanker hRM:

1. Cells stored in Bambanker grew faster during the next 28 days after thawing
2. The cell density was 96.6% when compared to the non-preserved control
3. The hexagonal shape and a monolayer of the cells was observed after thawing after 28 days of cultivation.
4. Cells stored with Bambanker showed key markers of the HCEC, comparable to the unfrozen cells ( ZO-1, N-cadherrin and Na+/Ka+-ATPase).
5. No fibroblastic changes were observed after cryopreservation.

Here is their comment:

“[…]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. […]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.”

Fig. 2 (A) from Feasibility of a cryopreservation of cultured human corneal endothelial cells
Okumura N, Kagami T, Watanabe K, Kadoya S, Sato M, et al. (2019) Feasibility of a cryopreservation of cultured human corneal endothelial cells. PLOS ONE 14(6):  e0218431.

https://doi.org/10.1371/journal.pone.0218431

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Bambanker hRM