Transplantation of human corneal limbal epithelial cell sheet harvested on synthesized carboxymethyl cellulose and dopamine in a limbal stem cell deficiency
Hun Lee
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Hun Lee and Jae H. Lee contributed equally to this study.Search for more papers by this authorJae Hyuck Lee
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Hun Lee and Jae H. Lee contributed equally to this study.Search for more papers by this authorSoyoung Hong
Biomedical Engineering Research Center, Asan Institute for Life Sciences, Asan Medical Center, Seoul, South Korea
Search for more papers by this authorJeong Hye Sunwoo
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Search for more papers by this authorHyun Tae Kim
Department of Ophthalmology, Chungbuk National University Hospital, Chungbuk National University College of Medicine, Cheongju, South Korea
Search for more papers by this authorEun-Soon Kim
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Search for more papers by this authorJae Yong Kim
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Search for more papers by this authorCorresponding Author
Changmo Hwang
Biomedical Engineering Research Center, Asan Institute for Life Sciences, Asan Medical Center, Seoul, South Korea
Department of Convergence Medicine, University of Ulsan College of Medicine, Seoul, South Korea
Correspondence
Hungwon Tchah, Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine 88, Olympic-Ro 43-Gil, Songpa-Gu, Seoul 05505, South Korea.
Email: [email protected]
Changmo Hwang, Biomedical Engineering Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, 88, Olympic-Ro 43-Gil, Songpa-Gu, Seoul 05505, South Korea.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Hungwon Tchah
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Correspondence
Hungwon Tchah, Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine 88, Olympic-Ro 43-Gil, Songpa-Gu, Seoul 05505, South Korea.
Email: [email protected]
Changmo Hwang, Biomedical Engineering Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, 88, Olympic-Ro 43-Gil, Songpa-Gu, Seoul 05505, South Korea.
Email: [email protected]
Search for more papers by this authorHun Lee
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Hun Lee and Jae H. Lee contributed equally to this study.Search for more papers by this authorJae Hyuck Lee
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Hun Lee and Jae H. Lee contributed equally to this study.Search for more papers by this authorSoyoung Hong
Biomedical Engineering Research Center, Asan Institute for Life Sciences, Asan Medical Center, Seoul, South Korea
Search for more papers by this authorJeong Hye Sunwoo
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Search for more papers by this authorHyun Tae Kim
Department of Ophthalmology, Chungbuk National University Hospital, Chungbuk National University College of Medicine, Cheongju, South Korea
Search for more papers by this authorEun-Soon Kim
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Search for more papers by this authorJae Yong Kim
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Search for more papers by this authorCorresponding Author
Changmo Hwang
Biomedical Engineering Research Center, Asan Institute for Life Sciences, Asan Medical Center, Seoul, South Korea
Department of Convergence Medicine, University of Ulsan College of Medicine, Seoul, South Korea
Correspondence
Hungwon Tchah, Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine 88, Olympic-Ro 43-Gil, Songpa-Gu, Seoul 05505, South Korea.
Email: [email protected]
Changmo Hwang, Biomedical Engineering Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, 88, Olympic-Ro 43-Gil, Songpa-Gu, Seoul 05505, South Korea.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Hungwon Tchah
Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea
Correspondence
Hungwon Tchah, Department of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine 88, Olympic-Ro 43-Gil, Songpa-Gu, Seoul 05505, South Korea.
Email: [email protected]
Changmo Hwang, Biomedical Engineering Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, 88, Olympic-Ro 43-Gil, Songpa-Gu, Seoul 05505, South Korea.
Email: [email protected]
Search for more papers by this authorAbstract
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.
CONFLICT OF INTEREST
The authors have no proprietary or financial interest in the materials presented herein.
Supporting Information
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| term3159-sup-0001-fig_s1.tif1.6 MB | Supplementary Material |
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