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GA Silva, JC Stendahl, C Czeisler, TA Neideen, RC Claussen, JA Kessler, SI Stupp; Semi-Dissociated Mixed Retinal Cultures Grown on a Fibrous Scaffold Modified by Self-Assembling Molecules Containing L-lysine . Invest. Ophthalmol. Vis. Sci. 2002;43(13):722.
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Purpose: To investigate the properties of primary semi-dissociated mixed retinal cultures grown on a biodegradable scaffold composed of poly-L-lactic acid (PLLA) fibers coated with self-assembling L-lysine containing molecules. Methods: The culture procedure was a modification of Dhingra et. al. (2001) Int. J. Devl. Neurosci. 19, 533-540. Briefly, neural retinas from P0-P1 Sprague-Dawley rat pups were removed and mechanically triturated first with a Pasteur pipette followed by an 18G needle. The resulting suspension of dissociated cells and small cell clusters of un-dissociated retina were plated onto lysine modified fibers, bare fiber controls, and poly-D-lysine (PDL) coated control glass cover slips. The PLLA substrates were constructed by pressing 15 µm diameter fibers under different pressures ranging from 8-350 MPa. Results: Good cellular adherence and survival was observed on lysine modified fibers and PDL controls in vitro and following their fixation and staining at 7 DIV under optical and scanning electron microscopy (SEM). SEM showed cellular aggregates qualitatively similar to SEM studies of intact retina, including neurite growth and development. Diffusion measurements of glucose for the 350 MPa lysine modified PLLA scaffold were ∼30 mg/dl/hr, indicating good matrix diffusion properties. Conclusion: Semi-dissociated mixed retinal cultures survive and grow on lysine modified fibers. Similar preparations have been grafted as free cell suspensions into animal models of, and human patients with, retinal degenerative disorders. The versatile construction and chemical properties of the substrates in this work can potentially be explored as a novel delivery system to improve the clinical outcome of such grafts.
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