Abstract
Human corneal endothelial cells (HCEnCs) are responsible for maintaining the transparency of the cornea. Damaged or diseased HCEnCs may cause blindness. Replacement of the diseased cells with a healthy donor endothelium is the only currently available treatment. Tissue-engineering can serve as an alternative to conventional donor corneal transplantation. Due to the global shortage of donor corneas, a wide interest in the development of cultured graft substitutes and artificial corneas has increased. Availability of the old donor corneas is higher especially for research. Although it can be proposed as a valuable source for cell culture, its less proliferative capability emerges a challenge for the researchers. This article describes the use of hyaluronic acid (HA) in combination with Rho-kinase inhibitor (ROCK) Y-27632 for the cultivation of HCEnCs from older donor corneas (age > 60 years). Four conditions including and excluding HA + ROCK and its effect on early attachment rates and proliferation was studied on forty-eight corneas. It was observed that HCEnCs reach confluence within 10–15 days when cultured with HA + ROCK. This approach improves the efficiency of cell adhesion due to force attachment. HCEnCs from old donor corneas can be cultured using this method which may further lead to cell-based therapy for treating corneal endothelial dysfunction.
Highlights
One of the most common approaches for therapeutic treatment and Human corneal endothelial cells (HCEnCs) regeneration includes the use of Rho-Kinase (ROCK) inhibitor for the development of allogeneic ex vivo expanded HCEnCs for transplantation[5]
Cultivated HCEnCs derived from older donors have lower proliferative capability, a senescent cell phenotype, and cell morphology, which suggest less functional ability than those derived from younger donors[19]
If the HCEnCs can be cultured from old donor corneas, the availability of donor corneas for isolating endothelial cells will increase and it will overcome the overall shortage of donor corneas available for transplantation[20, 21]
Summary
One of the most common approaches for therapeutic treatment and HCEnCs regeneration includes the use of Rho-Kinase (ROCK) inhibitor for the development of allogeneic ex vivo expanded HCEnCs for transplantation[5]. Ex-vivo expansion using ROCK inhibitor may allow potential cell-based therapy. Reported studies on culturing HCEnCs have been performed on younger donor corneas[12]. Most of the old donor corneas are easy to obtain for research due to its endothelial cell density that is less than the threshold required for transplantation. If the HCEnCs from the older donors can be cultured the availability of the source will be much higher compared to the younger donor corneas. The paper highlights four different conditions to identify the role of HA and Rho kinase (ROCK inhibitor) for force adherence in culture of HCEnCs which may eventually lead to higher number of corneal endothelial sheets from older donor corneas, reducing the requirement of human corneal tissues globally
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