June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Transforming growth factor beta secretion by human embryonic stem cell derived retinal pigment epithelium
Author Affiliations & Notes
  • Hossein Nazari Khanamiri
    Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
    Ophthalmology, Rassoul Akram Hospital, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran
  • Danhong Zhu
    Pathology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
    Doheny Eye Institute, Los Angeles, CA
  • Christine Spee
    Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
    Doheny Eye Institute, Los Angeles, CA
  • Mark Humayun
    Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
    Doheny Eye Institute, Los Angeles, CA
  • David Hinton
    Pathology, Keck School of Medicine of the University of Southern California, Los Angeles, CA
    Doheny Eye Institute, Los Angeles, CA
  • Footnotes
    Commercial Relationships Hossein Nazari Khanamiri, None; Danhong Zhu, None; Christine Spee, None; Mark Humayun, Bausch & Lomb (F), Bausch & Lomb (C), Bausch & Lomb (P), Bausch & Lomb (R), Bausch & Lomb (S), Alcon (C), Alcon (R), Iridex (P), Iridex (R), Replenish (I), Replenish (C), Replenish (R), Replenish (S), Second Sight (F), Second Sight (I), Second Sight (C), Second Sight (P), Second Sight (R), Second Sight (S), Regenerative Patch Technologies (I), Regenerative Patch Technologies (C); David Hinton, RPT (I), RPT (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2208. doi:
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      Hossein Nazari Khanamiri, Danhong Zhu, Christine Spee, Mark Humayun, David Hinton; Transforming growth factor beta secretion by human embryonic stem cell derived retinal pigment epithelium. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2208.

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

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Abstract

Purpose: Transforming growth factor beta (TGF-β), produced by retinal pigment epithelium (RPE), is a major component of the subretinal immune suppressive microenvironment. Implantation of human embryonic stem cell derived RPE (hESC-RPE) is emerging as a new strategy for the replacement of lost RPE cells in age-related macular degeneration. It is important to know whether these implanted hESC-RPE cells can produce cytokines involved in subretinal immune privilege environment. In the current study, we analyzed the ability of polarized hESC-RPE to produce TGF-β1 and -β2.

Methods: The H9 human embryonic stem cell line was used for derivation of RPE cells. hESC-RPE polarization (trans epithelial resistance above 300 Ohm/cm2) was achieved by long-term growth on permeable inserts. Apical and basal production of active TGF-β1 and -β2 was compared by ELISA in the medium collected from the apical and basal compartments. Total TGF-β1 and -β2 were measured by ELISA after acid activation. The absolute amount of secreted TGF-β was normalized to cellular total protein. Intracellular localization of the TGF-β1 and -β2 was visualized by immunostaining and confocal microscopy.

Results: The TGF-β1 was hardly detectable in the medium of polarized hESC-RPE. The level of latent (inactive) TGF-β2 was higher in the medium of the basal side, compared to the apical side of the hESC-RPE cells. However, the level of active TGF-β2 was higher in the medium from the apical side than the medium from the basal side. Active TGF-β2 was the major form of TGF-β2 secreted from the apical side, reaching to about 50% of total apically secreted TGF-β2.

Conclusions: TGF-β2 is the main TGF isoform secreted by hESC-RPE. More active TGF-β2 is secreted from the apical side of the polarized hESC-RPE, while more latent TGF-β2 is secreted from the basal side. Apically secreted active TGF-β2 may play a role in providing an immunosuppressive microenvironment for the implanted hESC-RPE cells.

Keywords: 701 retinal pigment epithelium • 721 stem cells • 490 cytokines/chemokines  
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