title: Characterization of cells from patient-derived fibrovascular membranes in proliferative diabetic retinopathy.
abstract: OBJECTIVE
  Epiretinal fibrovascular membranes (FVMs) are a hallmark of proliferative diabetic retinopathy (PDR). Surgical removal of FVMs is often indicated to treat tractional retinal detachment. This potentially informative pathological tissue is usually disposed of after surgery without further examination. We developed a method for isolating and characterizing cells derived from FVMs and correlated their expression of specific markers in culture with that in tissue.
  
  
  METHODS
  FVMs were obtained from 11 patients with PDR during diabetic vitrectomy surgery and were analyzed with electron microscopy (EM), comparative genomic hybridization (CGH), immunohistochemistry, and/or digested with collagenase II for cell isolation and culture. Antibody arrays and enzyme-linked immunosorbent assay (ELISA) were used to profile secreted angiogenesis-related proteins in cell culture supernatants.
  
  
  RESULTS
  EM analysis of the FVMs showed abnormal vessels composed of endothelial cells with large nuclei and plasma membrane infoldings, loosely attached perivascular cells, and stromal cells. The cellular constituents of the FVMs lacked major chromosomal aberrations as shown with CGH. Cells derived from FVMs (C-FVMs) could be isolated and maintained in culture. The C-FVMs retained the expression of markers of cell identity in primary culture, which define specific cell populations including CD31-positive, alpha-smooth muscle actin-positive (SMA), and glial fibrillary acidic protein-positive (GFAP) cells. In primary culture, secretion of angiopoietin-1 and thrombospondin-1 was significantly decreased in culture conditions that resemble a diabetic environment in SMA-positive C-FVMs compared to human retinal pericytes derived from a non-diabetic donor.
  
  
  CONCLUSIONS
  C-FVMs obtained from individuals with PDR can be isolated, cultured, and profiled in vitro and may constitute a unique resource for the discovery of cell signaling mechanisms underlying PDR that extends beyond current animal and cell culture models.
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA ; Retina Service, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, MA.
authors.forename: Leo A
authors.initials: LA
authors.lastname: Kim
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA.
authors.forename: Lindsay L
authors.initials: LL
authors.lastname: Wong
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA.
authors.forename: Dhanesh S
authors.initials: DS
authors.lastname: Amarnani
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Bigger-Allen
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA.
authors.forename: Christina K
authors.initials: CK
authors.lastname: Marko
authors.affiliation: Retina Service, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, MA.
authors.forename: Dean
authors.initials: D
authors.lastname: Eliott
authors.affiliation: Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK.
authors.forename: Vinay A
authors.initials: VA
authors.lastname: Shah
authors.affiliation: C.S. Kubik Laboratory for Neuropathology, Massachusetts General Hospital, Boston, MA.
authors.forename: Declan
authors.initials: D
authors.lastname: McGuone
authors.affiliation: C.S. Kubik Laboratory for Neuropathology, Massachusetts General Hospital, Boston, MA.
authors.forename: Anat O
authors.initials: AO
authors.lastname: Stemmer-Rachamimov
authors.affiliation: Ocular Genomics Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA.
authors.forename: Xiaowu
authors.initials: X
authors.lastname: Gai
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA ; Department of Pathology, Harvard Medical School, Boston, MA.
authors.forename: Patricia A
authors.initials: PA
authors.lastname: D'Amore
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA.
authors.forename: Joseph F
authors.initials: JF
authors.lastname: Arboleda-Velasquez
mesh_terms: D000199:Actins / Q000378:metabolism; D000328:Adult; D042683:Angiopoietin-1 / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D055028:Comparative Genomic Hybridization; D003930:Diabetic Retinopathy / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D019773:Epiretinal Membrane / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D005260:Female; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: C541116:ACTA2 protein, human; C467630:ANGPT1 protein, human; D000199:Actins; D042683:Angiopoietin-1; D005904:Glial Fibrillary Acidic Protein; D019408:Platelet Endothelial Cell Adhesion Molecule-1