Viability of human corneal keratocytes during organ culture

Abstract

The viability of human corneal keratocytes was assessed during four weeks of 'closed system' organ culture at 31 degrees C. After 28 days of culturing, the entire keratocyte population was still alive and viable because all cells incorporated uridine; a parameter for RNA-synthesis. During the first 14 days, mitoses were found in the anterior half of the stroma (0.23% mitoses per 48 h), while only few keratocytes were able to divide at day 28 (0.01% mitoses per 48 h). Metabolic parameters revealed a progressing acidosis in the medium with oxygen and glucose depletion. Immunological measurements of keratan sulphate proteoglycan suggested that approximately 1% of the total content was lost during the period. In conclusion, our current organ culture technique can maintain a viable keratocyte population for four weeks; a viable stroma can be grafted within this period.

Assessment of anti-scarring therapies in ex vivo organ cultured rabbit corneas

“…(Møller-Pedersen and Møller, 1996; Richard et al) Air/liquid interface organ cultures have been shown to improve epithelial cell morphology and preserve stromal keratocytes for at least 4 weeks. (Castro-Combs et al, 2008) In addition, our ex vivo model system was able to reproduce key aspects of corneal scarring reported in vivo for rabbits.…”

Corneal endothelium after refractive surgery

“…Other studies approximate the cell loss rate at 0.3% to 0.5% per year. 17,18,20,21 Systemic and ocular diseases also adversely affect endothelial cell populations. Patients with type II diabetes have greater endothelial morphology changes, but the overall effect of diabetes on ECD is debated.…”

Evaluation of 2 ex vivo Bovine Cornea Storage Protocols for Drug Delivery Applications

“…Here, ophthalmologists immediately transfer excised tissue into organ culture (OC) media and incubate it at physiological temperatures until it is required [11, 12]. The practice has been able to extend corneal transplantation viability for over a month notably through preservation of endothelial and keratocyte function [13, 14]. …”