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The cultivation of human retinal pigment epithelial cells on Bombyx mori silk fibroin

By Audra Shadforth, Karina A. George, Anthony Kwan, Traian V. Chirila and Damien G. Harkin

Abstract

We have presently evaluated membranes prepared from Bombyx mori silk fibroin (BMSF), for their potential use as a prosthetic Bruch’s membrane and carrier substrate for human retinal pigment epithelial (RPE) cell transplantation. Porous BMSF membranes measuring 3 μm in thickness were prepared from aqueous solutions (3% w/v) containing poly(ethylene oxide) (0.09%). The permeability coefficient for membranes was between 3 and 9 × 10-5 cm/s by using Allura red or 70 kDa FITC-dextran respectively. Average pore size (± sd) was 4.9 ± 2.3 µm and 2.9 ± 1.5 µm for upper and lower membrane surfaces respectively. Optimal attachment of ARPE-19 cells to BMSF membrane was achieved by pre-coating with vitronectin (1 µg/mL). ARPE-19 cultures maintained in low serum on BMSF membranes for approximately 8 weeks, developed a cobble-stoned morphology accompanied by a cortical distribution of F-actin and ZO-1. Similar results were obtained using primary cultures of human RPE cells, but cultures took noticeably longer to establish on BMSF compared with tissue culture plastic. These findings encourage further studies of BMSF as a substrate for RPE cell transplantation

Topics: 090301 Biomaterials, 111301 Ophthalmology, Silk fibroin, Retinal pigment epithelium, Bruch's membrane, Age-related macular degeneration, Transplantation
Publisher: Elsevier
Year: 2012
DOI identifier: 10.1016/j.biomaterials.2012.02.040
OAI identifier: oai:eprints.qut.edu.au:58570

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  1. A protocol for the culture and differentiation of highly polarized human retinal pigment epithelial cells.
  2. A tissue-engineered approach towards retinal repair: scaffolds for cell transplantation to the subretinal space.
  3. Age-related macular degeneration.
  4. (1996). ARPE-19, a human retinal pigment epithelial cell line with differentiated properties. Exp Eye Res
  5. (2008). Bombyx mori silk fibroin membranes as potential substrata for epithelial constructs used in the management of ocular surface disorders. Tissue Eng Part A
  6. (2009). Development of tissue-engineered membranes for the culture and transplantation of retinal pigment epithelial cells.
  7. Effects of culture conditions on heterogeneity and the apical junctional complex of the ARPE-19 cell line.
  8. Helicoidal multi-lamellar features of RGD-functionalized silk biomaterials for corneal tissue engineering.
  9. Human corneal endothelial cell growth on a silk fibroin membrane.
  10. Human corneal epithelial equivalents constructed on Bombyx mori silk fibroin membranes.
  11. Macromolecular diffusion characteristics of ageing human Bruch's membrane: implications for age-related macular degeneration (AMD).
  12. Novel role for DvE5-integrin in retinal adhesion and its diurnal peak.
  13. (2010). Porous Silk fibroin film as a transparent carrier for cultivated corneal epithelial sheets.
  14. RPE cell cultivation.
  15. Silk as a substratum for cell attachment and proliferation.
  16. Silk fibroin in ocular tissue reconstruction.
  17. Silk film biomaterials for cornea tissue engineering. Biomaterials 2009;30:1299-308.
  18. Silk-based biomaterials.
  19. Stem cell-based tissue engineering with silk biomaterials.
  20. (1999). The human retina and retinal pigment epithelium are abundant sources of vitronectin mRNA.
  21. The retinal pigment epithelium in visual function.
  22. Thin collagen film scaffolds for retinal epithelial cell culture.
  23. Vitronectin gene expression in the adult human retina. Invest Ophthalmol Vis Sci 1999;40:3305-

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