Abstract
Purpose :
Trying to improve the very limited proliferative ability of human corneal endothelial cells (HCECs) has remained a big challenge to researchers.In this study we investigated whether human adipose-derived stem cell–conditioned medium (ASC-CM) promote the proliferation and repair capacity of HCECs.
Methods :
Primary cultures of HCECs were purified following the method by stripping off Descemet’s membrane. The cells were cultivated in human corneal endothelium medium containing ASC-CM. Then the proliferative and repair capacity of HCECs were studied by cell proliferation assay and animal experiments. We created the corneal endothelial dysfunction models of rabbit by scraping the corneal endothelium. The cultured HCECs were injected into the anterior chamber，after which the changes of cornea were observed and assessed by slit lamp microscope, optical coherence tomography and in vivo confocal microscope. Immunofluorescence and histological examinations of the cornea tissues were performed either.
Results :
The cultivated HCECs could maintain fairly good proliferative and cell-based therapy capacity over 10 passages in this study.CEC relative markers such as zonula occludens-1 (ZO-1), Na+ /K+ ATPase and N-cadherin expressed in prolonged HCECs by Real-time reverse transcription–polymerase chain reaction (RT-PCR), Western Blot and immunofluorescence. Corneal transparence was achieved after the injection of cultivated HCECs which demonstrates the repair capacity of the cells.
Conclusions :
Using of ASC-CM not only stimulates the proliferation of HCECs in vitro but also promotes their repair capacity effectively. Thus more useful cells would be available for the research of HCECs and cell-based therapy for corneal endothelial dysfunction.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.