Reconstruction of Functional Ocular Surface by Acellular Porcine Cornea Matrix Scaffold and Limbal Stem Cells Derived from Human Embryonic Stem Cells

Abstract

Limbal stem cells (LSCs) derived from human embryonic stem cells (hESCs) hold great potential for cell-based therapies for ocular surface diseases. The cell-based therapies mainly depend on an appropriate differentiation proposal with a high efficiency and on a suitable scaffold. In this study, we aimed to establish a feasible and efficient strategy for inducing hESCs into LSC-like cells by the LSC conditioned medium. The induced cells possessed the similar morphologic characteristics and expression of normal LSCs and showed a strong clonogenic and proliferative capacity in vitro. To construct a tissue-engineering corneal graft, these differentiated cells were seeded on an acelluar porcine corneal matrix (APCM), which maintained the corneal basement membrane in vitro. After 14 days culture, these induced cells gave rise to stratified epithelial cell sheets on the APCM and the basal cells still kept LSC characteristics. In rabbit total limbal stem cell deficiency (LSCD) models, the tissue-engineering graft had the potential to reconstruct the damaged ocular surface and alleviated the invasion of corneal neovascularization. These findings indicated that the engraftment constructed with the APCM scaffold and hESC-derived LSCs might be a potential therapy option for ocular surface regeneration in LSCD cases.

Lamellar Keratoplasty Treatment of Fungal Corneal Ulcers With Acellular Porcine Corneal Stroma

“…The mesh structure in the matrix layer can provide space for the growth of mesenchymal cells and the exchange of metabolites. In addition, APCSs can also be used as seed carriers (21). Our previous animal study (13), through implantation of APCSs into rabbit corneal pockets and mouse subcutaneous tissue, showed that the APCS is unable to stimulate a host response in the cornea.…”

The Use of Stem Cells in Plastic and Reconstructive Surgery

“…With the use of specific media and growth factors, ESCs have been cultured in vitro into other cell types, including skeletal muscle, endothelial cells, chondrocytes and myocardiocytes, among others [3,4]. In later experiments on these stem cells, researchers have produced eyeball and vocal cord structures [5,6].…”

Biomaterials for Regenerative Medicine Approaches for the Anterior Segment of the Eye

“…Having produced the decellularized cornea there is then a choice of which cells to use to repopulate the construct for transplantation onto the injured cornea to promote the repair. [24,25] For example, human embryonic stem cells have been differentiated into LSC-like cells and used to repopulate acellular pig corneal matrix and following transplantation in a rabbit LSCD model was able to reconstruct the surface of the cornea and maintain the cornea/conjunctiva boundary. [26] Epithelial cells have also been isolated from human AM and seeded into decellularized pig cornea and shown to stratify after 7 d. When transplanted into a rabbit model it was found to integrate well into the host tissue and maintain corneal clarity.…”