title: Inhibition of TGF-β signaling enables human corneal endothelial cell expansion in vitro for use in regenerative medicine.
abstract: Corneal endothelial dysfunctions occurring in patients with Fuchs' endothelial corneal dystrophy, pseudoexfoliation syndrome, corneal endotheliitis, and surgically induced corneal endothelial damage cause blindness due to the loss of endothelial function that maintains corneal transparency. Transplantation of cultivated corneal endothelial cells (CECs) has been researched to repair endothelial dysfunction in animal models, though the in vitro expansion of human CECs (HCECs) is a pivotal practical issue. In this study we established an optimum condition for the cultivation of HCECs. When exposed to culture conditions, both primate and human CECs showed two distinct phenotypes: contact-inhibited polygonal monolayer and fibroblastic phenotypes. The use of SB431542, a selective inhibitor of the transforming growth factor-beta (TGF-β) receptor, counteracted the fibroblastic phenotypes to the normal contact-inhibited monolayer, and these polygonal cells maintained endothelial physiological functions. Expression of ZO-1 and Na(+)/K(+)-ATPase maintained their subcellular localization at the plasma membrane. Furthermore, expression of type I collagen and fibronectin was greatly reduced. This present study may prove to be the substantial protocol to provide the efficient in vitro expansion of HCECs with an inhibitor to the TGF-β receptor, and may ultimately provide clinicians with a new therapeutic modality in regenerative medicine for the treatment of corneal endothelial dysfunctions.
authors: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan.
authors: Naoki
authors: N
authors: Okumura
authors: EunDuck P
authors: EP
authors: Kay
authors: Makiko
authors: M
authors: Nakahara
authors: Junji
authors: J
authors: Hamuro
authors: Shigeru
authors: S
authors: Kinoshita
authors: Noriko
authors: N
authors: Koizumi
mesh_terms: D000818:Animals; D055419:Bone Morphogenetic Protein 7 / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism; D003315:Cornea / Q000378:metabolism* / Q000502:physiology; D042783:Endothelial Cells / Q000378:metabolism*; D004728:Endothelium, Corneal / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism / Q000502:physiology; D006801:Humans; D010641:Phenotype; D011323:Primates; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism; D044968:Regenerative Medicine / Q000379:methods; D015398:Signal Transduction; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014945:Wound Healing / Q000502:physiology*; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: D055419:Bone Morphogenetic Protein 7; D024042:Collagen Type I; D018125:Receptors, Transforming Growth Factor beta; C569042:TJP1 protein, human; D016212:Transforming Growth Factor beta; D062826:Zonula Occludens-1 Protein; D000254:Sodium-Potassium-Exchanging ATPase