title: Cultivation of corneal endothelial cells on a pericellular matrix prepared from human decidua-derived mesenchymal cells.
abstract: The barrier and pump functions of the corneal endothelium are essential for the maintenance of corneal transparency. Although corneal transplantation is the only current therapy for treating corneal endothelial dysfunction, the potential of tissue-engineering techniques to provide highly efficient and less invasive therapy in comparison to corneal transplantation has been highly anticipated. However, culturing human corneal endothelial cells (HCECs) is technically difficult, and there is no established culture protocol. The aim of this study was to investigate the feasibility of using a pericellular matrix prepared from human decidua-derived mesenchymal cells (PCM-DM) as an animal-free substrate for HCEC culture for future clinical applications. PCM-DM enhanced the adhesion of monkey CECs (MCECs) via integrin, promoted cell proliferation, and suppressed apoptosis. The HCECs cultured on the PCM-DM showed a hexagonal morphology and a staining profile characteristic of Na⁺/K⁺-ATPase and ZO-1 at the plasma membrane in vivo, whereas the control HCECs showed a fibroblastic phenotype. The cell density of the cultured HCECs on the PCM-DM was significantly higher than that of the control cells. These results indicate that PCM-DM provides a feasible xeno-free matrix substrate and that it offers a viable in vitro expansion protocol for HCECs while maintaining cellular functions for use as a subsequent clinical intervention for tissue-engineered based therapy of corneal endothelial dysfunction.
authors: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan.
authors: Ryohei
authors: R
authors: Numata
authors: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan ; Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors: Naoki
authors: N
authors: Okumura
authors: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan.
authors: Makiko
authors: M
authors: Nakahara
authors: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors: Morio
authors: M
authors: Ueno
authors: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors: Shigeru
authors: S
authors: Kinoshita
authors: Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
authors: Daisuke
authors: D
authors: Kanematsu
authors: Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
authors: Yonehiro
authors: Y
authors: Kanemura
authors: RIKEN Center for Developmental Biology, Hyogo, Japan.
authors: Yoshiki
authors: Y
authors: Sasai
authors: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan.
authors: Noriko
authors: N
authors: Koizumi
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D003656:Decidua / Q000166:cytology*; D004728:Endothelium, Corneal / Q000166:cytology*; D005260:Female; D006801:Humans; D008252:Macaca fascicularis; D059630:Mesenchymal Stem Cells / Q000737:chemistry*; D008875:Middle Aged; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*