May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Differences in differentiation potential between human retinal pigment epithelial cell cultures derived from fetal and nonfetal donors
Author Affiliations & Notes
  • T.R. Kinnick
    Physiological Sciences Program, Jules Stein Eye Institute,
    University of Arizona, Tucson, AZ
  • B. McKay
    Department of Ophthalmology, Department of Neurobiology,
    Cell Biology and Anatomy, Brain Research Institute,
    University of Arizona, Tucson, AZ
  • J. Hu
    Physiological Sciences Program, Jules Stein Eye Institute,
    University of California, Los Angeles, CA
  • D. Bok
    Department of Ophthalmology, Department of Neurobiology,
    Cell Biology and Anatomy, Brain Research Institute,
    University of California, Los Angeles, CA
  • A.D. Marmorstein
    Physiological Sciences Program, Jules Stein Eye Institute,
    Department of Ophthalmology, Department of Neurobiology,
    University of Arizona, Tucson, AZ
  • Footnotes
    Commercial Relationships  T.R. Kinnick, None; B. McKay, None; J. Hu, None; D. Bok, None; A.D. Marmorstein, None.
  • Footnotes
    Support  NIH Grant EY013160; NIH Grant EY014403
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 3672. doi:
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      T.R. Kinnick, B. McKay, J. Hu, D. Bok, A.D. Marmorstein; Differences in differentiation potential between human retinal pigment epithelial cell cultures derived from fetal and nonfetal donors . Invest. Ophthalmol. Vis. Sci. 2004;45(13):3672.

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

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Abstract

Abstract: : Purpose:To characterize the differences between retinal pigment epithelium (RPE) cell cultures derived from donors ranging in age from 20 wks gestation through 76 years. Methods: RPE cells were harvested and maintained according to published methods (Hu and Bok, Mol Vis 7: 14–9, 2000; McKay, ARVO abstract #387, 2003) which incorporate a calcium switch. Pigmentation of cultures was observed and documented qualitatively. Transepithelial resistances (TERs) were measured directly from Millicell HA inserts (Millipore Corporation, Bedford, MA) using an EVOM Epithelial Voltohmmeter. Protein markers (CRALBP, bestrophin) of RPE differentiation were assessed by Western blot. Results: Using modified Chee's medium with low calcium, only cells derived from fetal donors exhibited satisfactory levels of proliferation without prior enzymatic dissociation. Cultures derived from donors of all ages could be induced to pigment via calcium switch. However, only cultures derived from fetal donors expressed bestrophin and exhibited a significant TER when cultured on porous supports. Conclusions: Our findings indicate that RPE pigmentation is stimulated by Ca–dependent cell adhesion, however, only RPE cells derived from fetal donors could be induced to exhibit properties of later stage RPE differentiation –– including a high TER and expression of bestrophin.

Keywords: retinal pigment epithelium • calcium 
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