title: Matrigel and Activin A promote cell-cell contact and anti-apoptotic activity in cultured human retinal pigment epithelium cells.
abstract: Age-related macular degeneration (AMD) is a leading cause of blindness among the aging population. Currently, replacement of diseased retinal pigment epithelium (RPE) cells with transplanted healthy RPE cells could be a feasible approach for AMD therapy. However, maintaining cell-cell contact and good viability of RPE cells cultured in vitro is difficult and fundamentally determines the success of RPE cell transplantation. This study was conducted to examine the role of Matrigel and Activin A (MA) in regulating cell-cell contact and anti-apoptotic activity in human RPE (hRPE) cells, as assessed by atomic force microscopy (AFM), scanning electron microscope (SEM), immunofluorescence staining, quantitative polymerase chain reaction (qPCR) analysis, Annexin V/propidium iodide (PI) analysis, mitochondrial membrane potential (△Ψ m) assays, intracellular reactive oxygen species (ROS) assays and Western blotting. hRPE cells cultured in vitro could maintain their epithelioid morphology after MA treatment over at least 4 passages. The contact of N-cadherin to the lateral cell border was promoted in hRPE cells at P2 by MA. MA treatment also enhanced the expression of tight junction-associated genes and proteins, such as Claudin-1, Claudin-3, Occludin and ZO-1, as well as polarized ZO-1 protein distribution and barrier function, in cultured hRPE cells. Moreover, MA treatment decreased apoptotic cells, ROS and Bax and increased △Ψ m and Bcl2 in hRPE cells under serum withdrawal-induced apoptosis. In addition, MA treatment elevated the protein expression levels of β-catenin and its target proteins, including Cyclin D1, c-Myc and Survivin, as well as the gene expression levels of ZO-1, β-catenin, Survivin and TCF-4, all of which could be down-regulated by the Wnt/β-catenin pathway inhibitor XAV-939. Taken together, MA treatment could effectively promote cell-cell contact and anti-apoptotic activity in hRPE cells, partly involving the Wnt/β-catenin pathway. This study will benefit the understanding of hRPE cells and future cell therapy.
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou 510632, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Guo
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou 510632, China.
authors.forename: Deliang
authors.initials: D
authors.lastname: Zhu
authors.affiliation: The Department of Ophthalmology, the First Clinical Medical College, Jinan University, Guangzhou 510632, China.
authors.forename: Ruiling
authors.initials: R
authors.lastname: Lian
authors.affiliation: The Department of Ophthalmology, the First Clinical Medical College, Jinan University, Guangzhou 510632, China.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Han
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou 510632, China.
authors.forename: Yonglong
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Institute of Ophthalmology, Medical College, Jinan University, Jinan University, Guangzhou 510632, China.
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Li
authors.affiliation: Aier School of Ophthalmology, Central South University, Furong Middle Road 198#, Changsha 410015, China. Electronic address: tangsb@mail.sysu.edu.cn.
authors.forename: Shibo
authors.initials: S
authors.lastname: Tang
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou 510632, China; Institute of Ophthalmology, Medical College, Jinan University, Jinan University, Guangzhou 510632, China; The Department of Ophthalmology, the First Clinical Medical College, Jinan University, Guangzhou 510632, China. Electronic address: chenjiansu2000@163.com.
authors.forename: Jiansu
authors.initials: J
authors.lastname: Chen
mesh_terms: D028341:Activins / Q000494:pharmacology*; D022005:Adherens Junctions / Q000187:drug effects; D000328:Adult; D017304:Annexin A5 / Q000032:analysis; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D002478:Cells, Cultured; D057167:Claudins / Q000378:metabolism; D003094:Collagen / Q000494:pharmacology*; D004338:Drug Combinations; D004847:Epithelial Cells / Q000187:drug effects*; D005260:Female; D006801:Humans; D007797:Laminin / Q000494:pharmacology*; D008268:Macular Degeneration; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011419:Propidium / Q000032:analysis; D011509:Proteoglycans / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000187:drug effects
chemical_list: D017304:Annexin A5; D015820:Cadherins; D057167:Claudins; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D017382:Reactive Oxygen Species; C422405:activin A; D028341:Activins; C059485:matrigel; D011419:Propidium; D003094:Collagen
keywords: Activin A; Anti-apoptosis; Cell-cell contact; Human retinal pigmented epithelium; Matrigel