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S. Cheng, A. Kwan, Z. Barnard, Z. Zainuddin, D. Harkin, T. Chirila; Retinal Pigment Epithelial Cell Culture on Silk Substrate for Retinal Tissue Transplantation. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4278. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
A potential treatment for age-related macular degeneration is retinal pigment epithelial cell (RPE) transplantation. In this study we evaluated a novel substratum for RPE culture derived from domesticated silkworm Bombyx mori silk fibroin (BMSF), both alone or coated with a selection of extracellular matrix (ECM) proteins, and with or without serum.
The ARPE-19 cell line was seeded onto tissue culture plastic (TCP), BMSF membrane alone or BMSF membrane coated with laminin, vitronectin, fibronectin, a laminin-vitronectin-fibronectin combination (LVF) or collagen-IV. Samples were cultured under Foetal Bovine Serum (FBS) containing media for 72 hours, then fixed and nuclei stained with Hoechst and viable cells attached were counted. Experiments were repeated with serum starved ARPE-19 cells, which were seeded on to the different substrates, cultured under serum free conditions for 24 hours. Samples were fixed; nuclei stained and attached cells counted.
ARPE-19 cell growth on BMSF membrane demonstrated no statistical difference (P>0.05) to TCP under FBS containing culture condition. However, the ARPE-19 cell count on unmodified BMSF membrane was 60% of that on TCP under serum free condition (P<0.05). Cell count on ECM protein coated BMSF surpassed BMSF alone (vitronectin>collagen IV>fibronectin>LVF>laminin), except for laminin coated. Cell attachment on vitronectin or collagen IV coated BMSF was comparable to that seen in medium containing FBS on TCP.
ARPE-19 cells can be grown on BMSF membranes. Coating BMSF with ECM proteins increases attachment under serum free culture conditions. BMSF appears to be a potential substrate for RPE transplantation and merits further investigation.
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