June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Analysis and culture of cells from patient-derived membranes in proliferative diabetic retinopathy
Author Affiliations & Notes
  • Leo A Kim
    Retina Service, Massachusetts Eye and Ear Infirmary, Boston, MA
    Ophthalmology, Schepens Eye Research Institute / Mass Eye and Ear, Boston, MA
  • Lindsay L. Wong
    Ophthalmology, Schepens Eye Research Institute / Mass Eye and Ear, Boston, MA
  • Dhanesh Amarnani
    Ophthalmology, Schepens Eye Research Institute / Mass Eye and Ear, Boston, MA
  • Alex A. Bigger-Allen
    Ophthalmology, Schepens Eye Research Institute / Mass Eye and Ear, Boston, MA
  • Yang Hu
    Ophthalmology, Schepens Eye Research Institute / Mass Eye and Ear, Boston, MA
  • Christina Marko
    Ophthalmology, Schepens Eye Research Institute / Mass Eye and Ear, Boston, MA
  • Vinay Shah
    Ophthalmology, Dean McGee Eye Institute, Oklahoma City, OK
  • Dean Eliott
    Retina Service, Massachusetts Eye and Ear Infirmary, Boston, MA
  • Joseph F Arboleda-Velasquez
    Ophthalmology, Schepens Eye Research Institute / Mass Eye and Ear, Boston, MA
  • Patricia A D'Amore
    Ophthalmology, Schepens Eye Research Institute / Mass Eye and Ear, Boston, MA
  • Footnotes
    Commercial Relationships Leo Kim, None; Lindsay Wong, None; Dhanesh Amarnani, None; Alex Bigger-Allen, None; Yang Hu, None; Christina Marko, None; Vinay Shah, None; Dean Eliott, None; Joseph Arboleda-Velasquez, None; Patricia D'Amore, AGTC (C), Eleven Biotherapeutics (S)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 4683. doi:
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    • Get Citation

      Leo A Kim, Lindsay L. Wong, Dhanesh Amarnani, Alex A. Bigger-Allen, Yang Hu, Christina Marko, Vinay Shah, Dean Eliott, Joseph F Arboleda-Velasquez, Patricia A D'Amore; Analysis and culture of cells from patient-derived membranes in proliferative diabetic retinopathy. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4683.

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

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Abstract
 
Purpose
 

Formation of fibrovascular membranes (FVMs) is a hallmark of proliferative diabetic retinopathy (PDR). Complications of PDR such as retinal detachment and vitreous hemorrhage is addressed by surgical removal of FVMs. Here, we analyze and culture cells from surgically removed and patient-derived FVMs.

 
Methods
 

FVMs obtained from patients during vitrectomy surgery were analyzed by electron microscopy (EM), comparative genomic hybridization (CGH), immunohistology, and/or digested with collagenase II for cell isolation and culture. Antibody arrays and ELISA were used to profile angiogenesis-related proteins in cell culture supernatants.

 
Results
 

Ultrastructural analysis of FVM showed abnormal vessels within fibrous stroma. FVMs can be stratified by proliferative activity according to the distribution of specific cell markers. The cellular constituents of FVMs lacked major chromosomal aberrations as shown by CGH analysis. Cells derived from fibrovascular membranes could be isolated and maintained in culture. FVM cells can be maintained in primary culture and retain the expression of cell markers detected in FVM, which define specific cell populations including CD31, alpha-smooth muscle actin, and glial fibrillary acidic protein-positive cells. Secretion of angiopoietin-1 and thrombospondin-1 by these cultured cells was found to significantly decrease in high glucose and under hyperosmolar conditions compared to human retinal pericytes derived from a non-diabetic donor.

 
Conclusions
 

Cultured FVM cells recapitulate some of the functional characteristics of FVMs from patients with PDR by expressing characteristic cell markers found within FVMs, exhibiting cellular proliferation, and secreting angiogenic factors following unique patterns in response to environmental alterations. We believe these characteristics make cultured FVMs a useful tool in the evaluation of the underlying molecular mechanisms of PDR.  

 
(A) Pre-operative fundus photograph of the left eye shows FVM (dotted white line) involving the macula with fibrous tissue and aberrant blood vessels. (B) Post-operative fundus photograph 5 months post vitrectomy. (C) Bright field microscopy of FVM shows fibrous tissue and blood vessels (inset). (D) Bright field microscopy of cells derived from FVM reveals a morphologically heterogeneous mixture of fibroblastic and stellate cells. Scale bar = 100 µm.
 
(A) Pre-operative fundus photograph of the left eye shows FVM (dotted white line) involving the macula with fibrous tissue and aberrant blood vessels. (B) Post-operative fundus photograph 5 months post vitrectomy. (C) Bright field microscopy of FVM shows fibrous tissue and blood vessels (inset). (D) Bright field microscopy of cells derived from FVM reveals a morphologically heterogeneous mixture of fibroblastic and stellate cells. Scale bar = 100 µm.

 
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