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AE Fung, CJ Lee, T Leng, LV Bilbao, MC Peterman, MS Blumenkranz, SF Bent, HA Fishman; Tissue Engineered Lens Capsule as a Substrate for IPE and RPE Transplantation . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3452.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To explore the use of lens capsule with microprinted growth-controlling agents as a substrate for cultured IPE and RPE cells, as part of an effort to develop an autologous replacement for damaged RPE and Bruch’s membrane in patients with age-related macular degeneration (AMD). Methods: Photolithographic techniques were used to fabricate molds in photoresist on silicon substrates. Polydimethylsiloxane (PDMS) stamps for microcontact printing were made from these molds. The PDMS stamps were then used to wet-transfer a micropattern of cell-adhesion/spreading inhibitory molecules to the surface of human lens capsule which was coated on one side with a biodegradable matrix for stability. Iris pigment epithelium (IPE) harvested from rabbit with an enzymatic dissection technique were cultured in a standard IPE media (Ham F-12 with 20% FBS, gentamycin 50mg/ml) at 37 degrees Celsius. Human RPE (ARPE-19 cells) and rabbit IPE cells were grown on the lens capsule complexes, stained with anticytokeratin and anti-s100 antibodies to study their phenotypic characteristics, and examined with a combination of electron, confocal, fluorescence, and light microscopies. Results: Human lens capsule accepted microprinted patterns with a precision similar to that obtained on glass or synthetic polymers. RPE and IPE cells cultured onto untreated lens capsule showed spreading and formed into fusiform-appearing cells. In contrast, cells cultured on lens capsule with a hexagonal micropattern of inhibitory molecules retained epitheloid form. Conclusion: Inhibitory molecules can be microprinted onto human lens capsule in micropatterns to modulate the growth of RPE cells or IPE cells cultured on the surface. To our knowledge, this is the first time that microcontact printing has been successfully applied to lens capsule or human tissue. The ability to control the organization of RPE or IPE cell cultures and transplants may aid in the quest to treat age-related macular degeneration.
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