December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Semi-Dissociated Mixed Retinal Cultures Grown on a Fibrous Scaffold Modified by Self-Assembling Molecules Containing L-lysine
Author Affiliations & Notes
  • GA Silva
    Northwestern University Chicago IL
    Inst for Bioengineering and Nanoscience in Advanced Medicine
  • JC Stendahl
    Northwestern University Chicago IL
    Dept of Material Sciences and Engineering
  • C Czeisler
    Dept of Neurology
    Northwestern University Chicago IL
  • TA Neideen
    Northwestern University Chicago IL
    Inst for Bioengineering and Nanoscience in Advanced Medicine
  • RC Claussen
    Dept of Chemistry
    Northwestern University Chicago IL
  • JA Kessler
    Dept of Neurology
    Northwestern University Chicago IL
  • SI Stupp
    Northwestern University Chicago IL
    Dept of Material Sciences and Engineering
  • Footnotes
    Commercial Relationships   G.A. Silva, None; J.C. Stendahl, None; C. Czeisler, None; T.A. Neideen, None; R.C. Claussen, None; J.A. Kessler, None; S.I. Stupp, None. Grant Identification: NSF Grant DMR-0108342, NS Grant 20013, NS Grant 20778
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 722. doi:
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    • Get Citation

      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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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : 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.

Keywords: 560 retinal culture • 561 retinal degenerations: cell biology • 564 retinal development 
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