July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Gradual expansion of RPE-like cells in chick Müller glial culture
Author Affiliations & Notes
  • Run-Tao Yan
    Ophthalmology, Univ of Alabama at Birmingham, Birmingham, Alabama, United States
  • Shu-Zhen Wang
    Ophthalmology, Univ of Alabama at Birmingham, Birmingham, Alabama, United States
  • Footnotes
    Commercial Relationships   Run-Tao Yan, None; Shu-Zhen Wang, None
  • Footnotes
    Support  NIH/NEI grant EY011640, Research to Prevent Blindness, EyeSight Foundation of Alabama, and NIH/NEI core grant P30 EY003039
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 4577. doi:
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      Run-Tao Yan, Shu-Zhen Wang; Gradual expansion of RPE-like cells in chick Müller glial culture. Invest. Ophthalmol. Vis. Sci. 2018;59(9):4577.

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

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Purpose : Müller glia is considered to have the potential to give rise to multipotent progenitor cells. Previously we detected cells that were darkly-pigmented and positive for RPE65 (RPE-like) arising from Müller glial cell cultures established with embryonic day (E) 13 and E14 chick retina. To shed light on this presumed “Müller-to-RPE” phenomenon, this study examined whether it could occur when more developed retina was used and whether and how RPE-like cells amplified in the culture.

Methods : Primary Müller glial cell cultures were established with central and central-peripheral regions of chick retina of E16, when cell proliferation has ceased, and E18, when the retina becomes functional. The neural retina was separated from attached RPE by treatment with a Ca2+-free and Mg2+-free solution. Dissociated retinal cells were cultured at low density with infrequent medium replacement.

Results : After 7 days in vitro (DIV), Müller glial cells started to constitute the majority of the cell population. From 0 DIV to 25 DIV, the culture lacked cells containing the dark pigment granules typically present in RPE cells. By 28 DIV, darkly-pigmented cells, albert low in number, were visible under microscope, but not yet visible to the naked eyes. More cells became darkly-pigmented as the culture aged. Many of these cells localized/clustered as colonies visible to the naked eyes. Sequential photographs of the same culture over time showed that individual colonies of darkly-pigmented cells increased in their sizes (enlargement), but their pattern of distribution within the culture and their shapes (marked by darkly-pigmented cells) remained more or less the same throughout the period.

Conclusions : The emergence of darkly-pigmented cells in E18 culture suggests a Müller-to-RPE transition occurring in culture derived from well-developed (functional) retina. The conservation of the shape of melanized colonies and their overall distribution pattern implies a presence of localized factor(s) promoting RPE-like cell “recruiting.” It remains to be studied whether the localized factor(s) is of “progenitor proliferation” or of molecular cues guiding existing, neighboring cells to become RPE-like.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.


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