Abstract
Corneal disease affects 12.5 million individuals worldwide, with 2 million new cases each year. The standard treatment consists of a corneal transplantation from a human donor; however, the worldwide demand significantly exceeds the available supply. Lamellar endothelial keratoplasty, the replacement of only the endothelial layer of the cornea, can partially solve the problem. Progressive efforts have succeeded in expanding hCECs; however, the ability to expand hCECs is still limited, and new sources of CECs are being sought. Crucial advances have been achieved by the directed differentiation of embryonic or induced pluripotent stem cells, but these cells have disadvantages, such as the use of oncogenes, and are still difficult to establish. We aimed to transfer such knowledge to obtain hCECs from adipose tissue-derived adult mesenchymal stem cells (ADSC) by modifying four previously published procedures. We present several protocols capable of the directed differentiation of human ADSCs to hCECs. In our hands, the protocol by Ali et al. was the best adapted to such differentiation in terms of efficiency, time, and financial cost; however, the protocol by Wagoner et al. was the best for CEC marker expression. Our results broaden the type of cells of autologous extraocular origin that could be employed in the clinical setting for corneal endothelial deficiency.
Highlights
Corneal diseases are a major cause of vision loss, second only to cataracts in overall importance [1]
We aimed to transfer this knowledge to obtain human corneal endothelial cells (CEC) from human adipose tissue-derived mesenchymal stem cells, an easier to culture cell source, which is easy to obtain through elective liposuction, providing a high starting number of cells, with no viral or genetic manipulation necessary and, importantly, a possible autologous use
Typical CEC marker expression varies in actual human CECs with the culture, with high expression in directly isolated human CECs (d0, Figure 2C), decreasing in normal culturing conditions at passage 0 (d15, Figure 2C), and upregulating again after passaging at d43 at passage 1 (Figure 3D)
Summary
Corneal diseases are a major cause of vision loss, second only to cataracts in overall importance [1]. 12.5 million individuals worldwide experience unilateral or bilateral vision loss from corneal disease or injury [2], and approximately 2 million new cases are diagnosed each year [3]. The standard treatment for these conditions consists of a corneal transplantation (keratoplasty) from a human donor, whose success rate is approximately 80%, depending on the patient’s condition, with a
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