Potential Effect of Human Platelet Lysate on in vitro Expansion of Human Corneal Endothelial Cells Compared with Y-27632 ROCK Inhibitor.

Mohammad Amir Mishan,Zahra-Soheila Soheili,Sepehr Feizi,Mozhgan Rezaei Kanavi,Sahar Balagholi,Tahereh Chamani

doi:10.18502/jovr.v16i3.9431

Abstract

PurposeCorneal endothelial cell (CEC) therapy can be used as a promising therapeutic option for patients with various corneal endothelial dysfunctions. In this study, we compared the proliferative effect of human platelet lysate (HPL), as a xeno-free medium supplement, with Y-27632 Rho/rho-associated protein kinase (ROCK) inhibitor, as a well-known proliferative and adhesive agent for CECs, and fetal bovine serum (FBS) as the control, in the culture medium of human corneal endothelial cells (HCECs).MethodsWe isolated HCECs from human donors and treated the cells as three different treatment groups including 20% HPL only, 10 μM Y-27632 ROCK inhibitor, combination of 20% HPL and 10 μM Y-27632 ROCK inhibitor, and 20% FBS as the control group. ELISA cell proliferation assay and cell counting was performed on the treated cells. Finally, HCECs were characterized by morphology and immunocytochemistry (ICC).Results There was no significant proliferative effect of HPL on cell proliferation compared with the cells treated with Y-27632 ROCK inhibitor or the combination of HPL and Y-27632 ROCK inhibitor, but all the respected treatments had significant inducible effect on cell proliferation as compared with FBS-treated cells. The cells grown in all three treatment groups exhibited CEC morphology. Also, there was a higher expression of Na+/K+-ATPase and ZO-1, as CEC characteristic markers, in the culture of HCECs treated with HPL as compared with FBS.ConclusionHPL offers a xenofree and affordable medium supplement for CEC expansion that can be used in clinical applications.

Highlights

Corneal endothelium plays a pivotal role in the corneal transparency by regulating the flow of water from the aqueous humor into cornea.[1]
The proliferation of cultured human corneal endothelial cells (HCECs) after 24 hr treatment with human platelet lysate (HPL) was similar to those treated with Y-27632 rho-associated protein kinase (ROCK) inhibitor and with combination of HPL and Y-27632 ROCK inhibitor
The results of this study indicate that HCECs gained a potential proliferation after treatment with 20% HPL in comparison with fetal bovine serum (FBS) as the control group this proliferation effect was comparable to that in the Y-27632 ROCK inhibitortreated group

Summary

Introduction

Corneal endothelium plays a pivotal role in the corneal transparency by regulating the flow of water from the aqueous humor into cornea.[1] Corneal endothelial cell (CEC) density is above 2000 cells/mm in healthy people; corneal endothelial dysfunction and abnormal corneal hydration can occur following a decrease in CEC density to fewer than 400 cells/mm. [2] Loss of CECs occurs as the result of Fuchs’s endothelial corneal dystrophy (FECD) or following any form of corneal endothelial insult.[1] Current treatments for these situations include penetrating keratoplasty (PKP) and endothelial keratoplasty techniques for replacing the damaged endothelial layer.[3, 4] These surgical procedures are invasive and have several side effects. There is a need for an alternative solution based on cell therapy and/or tissue engineering to overcome the global shortage of donor corneas.[5, 6] cell therapy based on cultivated human corneal endothelial cells (HCECs) from cadaveric donor corneas has been suggested in several studies.[7, 8]

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