Structure and Barrier Properties of Human Embryonic Stem Cell–Derived Retinal Pigment Epithelial Cells Are Affected by Extracellular Matrix Protein Coating
| Autor: | Heidi Hongisto, Kai Kaarniranta, Hannu Uusitalo, Heli Skottman, Anni Sorkio, Kati Juuti-Uusitalo |
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| Rok vydání: | 2014 |
| Předmět: |
Cellular differentiation
Blotting Western Biomedical Engineering Bioengineering Retinal Pigment Epithelium Biochemistry Cell Line Biomaterials Extracellular matrix Coated Materials Biocompatible Laminin medicine Humans Embryonic Stem Cells Extracellular Matrix Proteins Retinal pigment epithelium biology Pigmentation Cell Differentiation Epithelial Cells Original Articles Embryonic stem cell eye diseases Cell biology Fibronectin medicine.anatomical_structure Cell culture biology.protein Basal lamina sense organs |
| Zdroj: | Tissue Engineering Part A. :140120073644000 |
| ISSN: | 1937-335X 1937-3341 |
| Popis: | Extracellular matrix (ECM) interactions play a vital role in cell morphology, migration, proliferation, and differentiation of cells. We investigated the role of ECM proteins on the structure and function of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells during their differentiation and maturation from hESCs into RPE cells in adherent differentiation cultures on several human ECM proteins found in native human Bruch's membrane, namely, collagen I, collagen IV, laminin, fibronectin, and vitronectin, as well as on commercial substrates of xeno-free CELLstart™ and Matrigel™. Cell pigmentation, expression of RPE-specific proteins, fine structure, as well as the production of basal lamina by hESC-RPE on different protein coatings were evaluated after 140 days of differentiation. The integrity of hESC-RPE epithelium and barrier properties on different coatings were investigated by measuring transepithelial resistance. All coatings supported the differentiation of hESC-RPE cells as demonstrated by early onset of cell pigmentation and further maturation to RPE monolayers after enrichment. Mature RPE phenotype was verified by RPE-specific gene and protein expression, correct epithelial polarization, and phagocytic activity. Significant differences were found in the degree of RPE cell pigmentation and tightness of epithelial barrier between different coatings. Further, the thickness of self-assembled basal lamina and secretion of the key ECM proteins found in the basement membrane of the native RPE varied between hESC-RPE cultured on compared protein coatings. In conclusion, this study shows that the cell culture substrate has a major effect on the structure and basal lamina production during the differentiation and maturation of hESC-RPE potentially influencing the success of cell integrations and survival after cell transplantation. |
| Databáze: | OpenAIRE |
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