Transplantation of human corneal limbal epithelial cell sheet harvested on synthesized carboxymethyl cellulose and dopamine in a limbal stem cell deficiency.

Abstract

This study aimed to evaluate the efficacy and safety of transplantation with human corneal limbal epithelial (HCLE) cell sheets cultured on carboxymethyl cellulose (CMC)-dopamine (DA)-coated substrates and harvested via enzymatic digestion of CMC with cellulase in a rabbit animal model of limbal stem cell deficiency (LSCD). Synthesized CMC-DA was pretreated onto the surface of culture plates. Then, HCLE cells were cultured on precoated CMC-DA and HCLE cell sheets were harvested using cellulase-containing cell culture medium. HCLE cell sheets were evaluated using a live/dead assay, histological examination, and immunofluorescence staining. For in vivo assessment, HCLE cell sheets were transplanted in a rabbit model of LSCD for 2 weeks to determine the effectiveness of the repair. Primary culture of HCLE cells stained positively for p63, cytokeratin (CK)15, and CK12. HCLE cell sheets were generated with a well-preserved morphology and transparency ranging in size from 15 to 19 mm after cellulase-assisted cell sheet generation. HCLE cell sheets uniformly stained positively for human mitochondria, p63, CK15, CK12, CK3/2p, and zonula occludens (ZO)-1. HCLE cell sheet transplantation in a rabbit model of LSCD improved the corneal opacity and neovascularization scores. Transplanted HCLE cell sheets stained positively for p63 and CK12. Transplantation of HCLE cell sheets harvested on CMC-DA coating combined with cellulase is a safe and efficient procedure for corneal epithelial regeneration in a rabbit model of LSCD. This system could enable a promising strategy to regenerate corneal epithelium by transplantation in ocular surface disorders.

• Authors: Hun Lee, Jae Hyuck Lee, Soyoung Hong, Jeong Hye Sunwoo, Hyun Tae Kim, Eun-Soon Kim, Jae Yong Kim, Changmo Hwang, Hungwon Tchah
• Journal: Journal of tissue engineering and regenerative medicine (J Tissue Eng Regen Med) Vol. 15 Issue 2 Pg. 139-149 (02 2021) ISSN: 1932-7005 [Electronic] England
• PMID: 33210832 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2020 John Wiley & Sons Ltd.

Chemical References

• Carboxymethylcellulose Sodium
• Dopamine

Topics

• Animals
• Carboxymethylcellulose Sodium (pharmacology)
• Cornea
• Corneal Diseases (metabolism, pathology, surgery)
• Disease Models, Animal
• Dopamine (pharmacology)
• Epithelial Cells (metabolism, pathology, transplantation)
• Epithelium, Corneal (metabolism, pathology, transplantation)
• Heterografts
• Humans
• Limbus Corneae (metabolism, pathology)
• Rabbits
• Stem Cells