PubTransformer

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PMID- 28777835
OWN - NLM
STAT- MEDLINE
DA  - 20170804
DCOM- 20170811
LR  - 20170811
IS  - 1552-5783 (Electronic)
IS  - 0146-0404 (Linking)
VI  - 58
IP  - 10
DP  - 2017 Aug 01
TI  - Effect of Methotrexate on an In Vitro Patient-Derived Model of Proliferative
      Vitreoretinopathy.
PG  - 3940-3949
LID - 10.1167/iovs.16-20912 [doi]
AB  - Purpose: The purpose of this study was to develop a method for isolating,
      culturing, and characterizing cells from patient-derived membranes in
      proliferative vitreoretinopathy (PVR) to be used for drug testing. Methods: PVR
      membranes were obtained from six patients with grade C PVR. Membrane fragments
      were analyzed by gross evaluation, fixed for immunohistologic studies to
      establish cell identity, or digested with collagenase II to obtain single cell
      suspensions for culture. PVR-derived primary cultures were used to examine the
      effects of methotrexate (MTX) on proliferation, migration, and cell death.
      Results: Gross analysis of PVR membranes showed presence of pigmented cells,
      indicative of retinal pigment epithelial cells. Immunohistochemistry identified
      cells expressing alpha-smooth muscle actin, glial fibrillary acidic protein,
      Bestrophin-1, and F4/80, suggesting the presence of multiple cell types in PVR.
      Robust PVR primary cultures (C-PVR) were successfully obtained from human
      membranes, and these cells retained the expression of cell identity markers in
      culture. C-PVR cultures formed membranes and band-like structures in culture
      reminiscent of the human condition. MTX significantly reduced the proliferation
      and band formation of C-PVR, whereas it had no significant effect on cell
      migration. MTX also induced regulated cell death within C-PVR as assessed by
      increased expression of caspase-3/7. Conclusions: PVR cells obtained from human
      membranes can be successfully isolated, cultured, and profiled in vitro. Using
      these primary cultures, we identified MTX as capable of significantly reducing
      growth and inducing cell death of PVR cells in vitro.
FAU - Amarnani, Dhanesh
AU  - Amarnani D
AD  - Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston,
      Massachusetts, United States.
FAU - Machuca-Parra, Arturo Israel
AU  - Machuca-Parra AI
AD  - Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston,
      Massachusetts, United States.
FAU - Wong, Lindsay L
AU  - Wong LL
AD  - Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston,
      Massachusetts, United States.
FAU - Marko, Christina K
AU  - Marko CK
AD  - Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston,
      Massachusetts, United States.
FAU - Stefater, James A
AU  - Stefater JA
AD  - Retina Service, Massachusetts Eye and Ear, Department of Ophthalmology, Harvard
      Medical School, Boston, Massachusetts, United States.
FAU - Stryjewski, Tomasz P
AU  - Stryjewski TP
AD  - Retina Service, Massachusetts Eye and Ear, Department of Ophthalmology, Harvard
      Medical School, Boston, Massachusetts, United States.
FAU - Eliott, Dean
AU  - Eliott D
AD  - Retina Service, Massachusetts Eye and Ear, Department of Ophthalmology, Harvard
      Medical School, Boston, Massachusetts, United States.
FAU - Arboleda-Velasquez, Joseph F
AU  - Arboleda-Velasquez JF
AD  - Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston,
      Massachusetts, United States.
FAU - Kim, Leo A
AU  - Kim LA
AD  - Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston,
      Massachusetts, United States.
AD  - Retina Service, Massachusetts Eye and Ear, Department of Ophthalmology, Harvard
      Medical School, Boston, Massachusetts, United States.
LA  - eng
GR  - K12 EY016335/EY/NEI NIH HHS/United States
GR  - P30 EY003790/EY/NEI NIH HHS/United States
GR  - R00 EY021624/EY/NEI NIH HHS/United States
GR  - R21 EY027061/EY/NEI NIH HHS/United States
PT  - Journal Article
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Biomarkers)
RN  - 0 (Extracellular Matrix Proteins)
RN  - 0 (Immunosuppressive Agents)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - YL5FZ2Y5U1 (Methotrexate)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Apoptosis/drug effects
MH  - Biomarkers/metabolism
MH  - Cell Culture Techniques
MH  - Cell Movement/physiology
MH  - Cell Proliferation/physiology
MH  - Cell Separation
MH  - Epiretinal Membrane/*drug therapy/metabolism/pathology
MH  - Extracellular Matrix Proteins/metabolism
MH  - Female
MH  - Fluorescent Antibody Technique, Indirect
MH  - Humans
MH  - Immunosuppressive Agents/*pharmacology
MH  - Male
MH  - Methotrexate/*pharmacology
MH  - Middle Aged
MH  - Models, Biological
MH  - Phenotype
MH  - Retinal Detachment/complications
MH  - Retinal Pigment Epithelium/*drug effects/metabolism/pathology
MH  - Tumor Necrosis Factor-alpha/pharmacology
MH  - Vitreoretinopathy, Proliferative/*drug therapy/etiology/metabolism/pathology
PMC - PMC5544356
EDAT- 2017/08/05 06:00
MHDA- 2017/08/12 06:00
CRDT- 2017/08/05 06:00
AID - 2647773 [pii]
AID - 10.1167/iovs.16-20912 [doi]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 2017 Aug 1;58(10):3940-3949. doi:
      10.1167/iovs.16-20912.