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Sugino IK, Sun Q, Springer C, et al. Two Bioactive Molecular Weight Fractions of a Conditioned Medium Enhance RPE Cell Survival on Age-Related Macular Degeneration and Aged Bruch's Membrane. Transl Vis Sci Technol. 2016;5(1):8doi: 10.1167/tvst.5.1.8.
Sugino, I. K., Sun, Q., Springer, C., Cheewatrakoolpong, N., Liu, T., Li, H., & Zarbin, M. A. (2016). Two Bioactive Molecular Weight Fractions of a Conditioned Medium Enhance RPE Cell Survival on Age-Related Macular Degeneration and Aged Bruch's Membrane. Translational vision science & technology, 5(1), 8. https://doi.org/10.1167/tvst.5.1.8
Sugino, Ilene K, et al. "Two Bioactive Molecular Weight Fractions of a Conditioned Medium Enhance RPE Cell Survival on Age-Related Macular Degeneration and Aged Bruch's Membrane." Translational vision science & technology vol. 5,1 (2016): 8. doi: https://doi.org/10.1167/tvst.5.1.8
Sugino IK, Sun Q, Springer C, Cheewatrakoolpong N, Liu T, Li H, Zarbin MA. Two Bioactive Molecular Weight Fractions of a Conditioned Medium Enhance RPE Cell Survival on Age-Related Macular Degeneration and Aged Bruch's Membrane. Transl Vis Sci Technol. 2016 Feb 22;5(1):8. doi: 10.1167/tvst.5.1.8. eCollection 2016 Feb. PMID: 26933521; PMCID: PMC4771074.
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Transl Vis Sci Technol. 2016 Feb 22;5(1):8. doi: 10.1167/tvst.5.1.8. eCollection 2016 Feb.

Two Bioactive Molecular Weight Fractions of a Conditioned Medium Enhance RPE Cell Survival on Age-Related Macular Degeneration and Aged Bruch's Membrane.

Translational vision science & technology

Ilene K Sugino, Qian Sun, Carola Springer, Noounanong Cheewatrakoolpong, Tong Liu, Hong Li, Marco A Zarbin

Affiliations

  1. Institute of Ophthalmology and Visual Science Rutgers, New Jersey Medical School, Newark, NJ, USA.
  2. Department of Biochemistry and Molecular Biology, Center for Advanced Proteomics Research, Neuroproteomics Core Facility, Rutgers, New Jersey Medical School, Newark, NJ, USA.

PMID: 26933521 PMCID: PMC4771074 DOI: 10.1167/tvst.5.1.8

Abstract

PURPOSE: To characterize molecular weight fractions of bovine corneal endothelial cell conditioned medium (CM) supporting retinal pigment epithelium (RPE) cell survival on aged and age-related macular degeneration (AMD) Bruch's membrane.

METHODS: CM was subject to size separation using centrifugal filters. Retentate and filtrate fractions were tested for bioactivity by analyzing RPE survival on submacular Bruch's membrane of aged and AMD donor eyes and behavior on collagen I-coated tissue culture wells. Protein and peptide composition of active fractions was determined by mass spectrometry.

RESULTS: Two bioactive fractions, 3-kDa filtrate and a 10-50-kDa fraction, were necessary for RPE survival on aged and AMD Bruch's membrane. The 3-kDa filtrate, but not the 10-50-kDa fraction, supported RPE growth on collagen 1-coated tissue culture plates. Mass spectrometry of the 10-50-kDa fraction identified 175 extracellular proteins, including growth factors and extracellular matrix molecules. Transforming growth factor (TGF)β-2 was identified as unique to active CM. Peptides representing 29 unique proteins were identified in the 3-KDa filtrate.

CONCLUSIONS: These results indicate there is a minimum of two bioactive molecules in CM, one found in the 3-kDa filtrate and one in the 10-50-kDa fraction, and that bioactive molecules in both fractions must be present to ensure RPE survival on Bruch's membrane. Mass spectrometry analysis suggested proteins to test in future studies to identify proteins that may contribute to CM bioactivity.

TRANSLATIONAL RELEVANCE: Results of this study are the first steps in development of an adjunct to cell-based therapy to ensure cell transplant survival and functionality in AMD patients.

Keywords: Bruch's membrane; age-related macular degeneration; cell survival; cell transplantation; retinal pigment epithelium

References

  1. Ophthalmic Res. 2000 Sep-Oct;32(5):244-8 - PubMed
  2. FEBS J. 2013 May;280(10 ):2120-37 - PubMed
  3. J Proteome Res. 2013 Jul 5;12(7):3496-510 - PubMed
  4. Exp Eye Res. 2007 Mar;84(3):464-72 - PubMed
  5. Exp Eye Res. 2013 Oct;115:13-21 - PubMed
  6. Graefes Arch Clin Exp Ophthalmol. 2012 Jan;250(1):15-23 - PubMed
  7. Invest Ophthalmol Vis Sci. 2010 Feb;51(2):1190-7 - PubMed
  8. Stem Cell Reports. 2015 May 12;4(5):860-72 - PubMed
  9. J Cell Sci. 2002 Nov 15;115(Pt 22):4201-14 - PubMed
  10. J Cell Physiol. 2005 Apr;203(1):35-43 - PubMed
  11. Invest Ophthalmol Vis Sci. 2009 Apr;50(4):1903-10 - PubMed
  12. Stem Cells. 2010 Apr;28(4):810-20 - PubMed
  13. Invest Ophthalmol Vis Sci. 1990 May;31(5):839-46 - PubMed
  14. Exp Eye Res. 2006 Jul;83(1):212-22 - PubMed
  15. Exp Eye Res. 2001 Oct;73(4):509-20 - PubMed
  16. Curr Eye Res. 2000 Jun;20(6):480-7 - PubMed
  17. Mol Vis. 2008 Jul 30;14:1358-72 - PubMed
  18. Mol Cell Proteomics. 2008 Jul;7(7):1349-61 - PubMed
  19. Arch Biochem Biophys. 2013 Jun;534(1-2):71-87 - PubMed
  20. Arch Ophthalmol. 1990 Mar;108(3):421-9 - PubMed
  21. Trans Am Ophthalmol Soc. 2004;102:123-37; discussion 137-8 - PubMed
  22. Invest Ophthalmol Vis Sci. 2011 Jul 07;52(8):5068-78 - PubMed
  23. Int J Ophthalmol. 2013;6(1):8-14 - PubMed
  24. Cytokine Growth Factor Rev. 2006 Aug;17(4):295-304 - PubMed
  25. Exp Eye Res. 2007 Dec;85(6):762-71 - PubMed
  26. Graefes Arch Clin Exp Ophthalmol. 1994 Mar;232(3):133-40 - PubMed
  27. Curr Eye Res. 1990 Apr;9(4):323-35 - PubMed
  28. J Clin Invest. 2011 Aug;121(8):3005-23 - PubMed
  29. Lancet. 2015 Feb 7;385(9967):509-16 - PubMed
  30. Histol Histopathol. 2003 Oct;18(4):1275-88 - PubMed
  31. Fibrogenesis Tissue Repair. 2012 Jun 06;5(Suppl 1):S24 - PubMed
  32. Invest Ophthalmol Vis Sci. 2011 Jul 01;52(8):4979-97 - PubMed
  33. Invest Ophthalmol Vis Sci. 2007 Dec;48(12):5722-32 - PubMed
  34. Exp Eye Res. 2005 Nov;81(5):581-91 - PubMed
  35. Invest Ophthalmol Vis Sci. 2014 Feb 26;55(2):1191-202 - PubMed
  36. Invest Ophthalmol Vis Sci. 2013 Nov 08;54(12):7329-37 - PubMed
  37. Trans Am Ophthalmol Soc. 2003;101:499-520 - PubMed
  38. Lab Invest. 2005 Jul;85(7):838-50 - PubMed
  39. FEBS J. 2010 Oct;277(19):3864-75 - PubMed
  40. Invest Ophthalmol Vis Sci. 2011 Mar 10;52(3):1345-58 - PubMed
  41. J Biol Chem. 1999 Aug 6;274(32):22755-62 - PubMed
  42. Stem Cells Int. 2012;2012:123030 - PubMed
  43. Matrix Biol. 2008 Mar;27(2):150-60 - PubMed
  44. Acta Biomater. 2011 Feb;7(2):463-77 - PubMed
  45. Curr Eye Res. 2009 Oct;34(10):867-76 - PubMed
  46. Tsitologiia. 2012;54(11):823-30 - PubMed
  47. Exp Eye Res. 2003 Nov;77(5):593-9 - PubMed
  48. FEBS J. 2006 Dec;273(23 ):5231-44 - PubMed
  49. Biochim Biophys Acta. 2006 Jan;1765(1):25-37 - PubMed
  50. Br J Ophthalmol. 2006 Aug;90(8):1052-9 - PubMed
  51. Invest Ophthalmol Vis Sci. 2011 Dec 20;52(13):9598-609 - PubMed
  52. Neurosci Lett. 2013 Aug 26;548:249-54 - PubMed
  53. Invest Ophthalmol Vis Sci. 2008 Sep;49(9):4078-88 - PubMed
  54. Growth Factors. 2008 Apr;26(2):80-91 - PubMed
  55. Toxicol In Vitro. 2010 Jun;24(4):1053-63 - PubMed

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