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Updated: Apr 8, 2026
An Ex Vivo Tissue Culture Model for Fibrovascular Complications in Proliferative Diabetic Retinopathy
Published on: January 25, 2019
Characterization of cells from patient-derived fibrovascular membranes in proliferative diabetic retinopathy
Leo A Kim1, Lindsay L Wong2, Dhanesh S Amarnani2
1Schepens 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.
Purpose:
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.
Insights
Researchers isolated and cultured cells from fibrovascular membranes (FVMs) in proliferative diabetic retinopathy (PDR). These cells offer a new model for studying PDR mechanisms beyond traditional methods.
Area of Science:
- Ophthalmology
- Cell Biology
- Diabetic Retinopathy Research
Background:
- Epiretinal fibrovascular membranes (FVMs) are characteristic of proliferative diabetic retinopathy (PDR).
- Surgical removal of FVMs is common for treating tractional retinal detachment.
- This pathological tissue is typically discarded without further analysis.
Purpose of the Study:
- To develop a method for isolating and characterizing cells from FVMs.
- To correlate the expression of specific markers in cultured FVM cells with their expression in tissue.
- To establish an in vitro model for studying PDR.
Main Methods:
- FVMs were obtained from 11 PDR patients during vitrectomy.
- Techniques included electron microscopy (EM), comparative genomic hybridization (CGH), and immunohistochemistry.
- Cells were isolated and cultured, with secreted proteins analyzed by antibody arrays and ELISA.
Main Results:
- EM revealed abnormal vessels with specific endothelial, perivascular, and stromal cells.
- CGH indicated a lack of major chromosomal aberrations in FVM cellular constituents.
- Cultured FVM cells (C-FVMs) retained identity markers (CD31, SMA, GFAP).
- SMA-positive C-FVMs showed decreased secretion of angiopoietin-1 and thrombospondin-1 in diabetic-like conditions compared to non-diabetic pericytes.
Conclusions:
- Cells derived from PDR FVMs (C-FVMs) can be successfully isolated and cultured.
- C-FVMs provide a valuable in vitro resource for discovering cell signaling mechanisms in PDR.
- This model offers advantages over existing animal and cell culture models for PDR research.
