PMID- 11818403
OWN - NLM
STAT- MEDLINE
DCOM- 20020214
LR  - 20061115
IS  - 0146-0404 (Print)
IS  - 0146-0404 (Linking)
VI  - 43
IP  - 2
DP  - 2002 Feb
TI  - Characterization of genetically modified human retinal pigment epithelial cells 
      developed for in vitro and transplantation studies.
PG  - 546-55
AB  - PURPOSE: To develop, by specific genetic modification, a differentiated human 
      retinal pigment epithelial (RPE) cell line with an extended life span that can be 
      used for investigating their function in vitro and for in vivo transplantation 
      studies. METHODS: Primary human RPE cells were genetically modified by 
      transfecting with a plasmid encoding the simian virus (SV)40 large T antigen. 
      After characterization, two cell lines, designated h1RPE-7 and h1RPE-116, were 
      chosen for further investigation, along with the spontaneously derived RPE cell 
      line ARPE-19. Factors reported to be important in RPE and photoreceptor cell 
      function and survival in vivo were examined. RESULTS: Both h1RPE-7 and h1RPE-116 
      cells exhibited epithelial morphology, expressed cytokeratins, and displayed 
      junctional distribution of ZO-1, p100-p120 and beta-catenin. The cells expressed 
      mRNA for RPE65 and cellular retinaldehyde-binding protein (CRALBP) and the 
      trophic and growth factors brain-derived neurotropic factor (BDNF), ciliary 
      neurotrophic factor (CNTF), basic fibroblast growth factor (bFGF), pigment 
      epithelium-derived factor (PEDF), nerve growth factor (NGF), platelet-derived 
      growth factor (PDGF)-alpha, insulin-like growth factor (IGF)-1, and vascular 
      endothelial growth factor (VEGF). Secreted BDNF, bFGF, and VEGF, but not CNTF, 
      were identified in cell supernatants. The cell lines constitutively expressed 
      HLA-ABC, CD54, CD58, and CD59. After activation with IFN-gamma both HLA-ABC and 
      CD54 were upregulated, and the expression of HLA-DR was induced. Both cell lines 
      failed to express CD80, CD86, CD40, or CD48 in vitro and in a mixed lymphocyte 
      reaction were unable to induce T-cell proliferation. Fas ligand (CD95L) was not 
      detected in vitro by RT-PCR. Similar results were obtained with the ARPE-19 cell 
      line. CONCLUSIONS: RPE lines h1RPE-7 and h1RPE-116 retain many of the morphologic 
      and biochemical characteristics of RPE cells in vivo and may serve as a source of 
      cells for in vitro analysis of RPE cell function, as well as for orthotopic 
      transplantation studies.
FAU - Kanuga, Naheed
AU  - Kanuga N
AD  - Division of Cell Biology, Institute of Ophthalmology, University College London, 
      London, United Kingdom.
FAU - Winton, Helen L
AU  - Winton HL
FAU - Beauchene, Laurence
AU  - Beauchene L
FAU - Koman, Ahmet
AU  - Koman A
FAU - Zerbib, Anne
AU  - Zerbib A
FAU - Halford, Stephanie
AU  - Halford S
FAU - Couraud, Pierre-Olivier
AU  - Couraud PO
FAU - Keegan, David
AU  - Keegan D
FAU - Coffey, Pete
AU  - Coffey P
FAU - Lund, Raymond D
AU  - Lund RD
FAU - Adamson, Peter
AU  - Adamson P
FAU - Greenwood, John
AU  - Greenwood J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Antigens, Polyomavirus Transforming)
RN  - 0 (DNA Primers)
RN  - 0 (Eye Proteins)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Antigens, Polyomavirus Transforming/genetics
MH  - Cell Line, Transformed
MH  - Cell Separation
MH  - Cell Survival
MH  - Cell Transplantation
MH  - DNA Primers/chemistry
MH  - Eye Proteins/genetics/metabolism
MH  - Female
MH  - Fluorescent Antibody Technique, Indirect
MH  - Humans
MH  - Lymphocyte Activation
MH  - Middle Aged
MH  - Pigment Epithelium of Eye/*cytology/metabolism/transplantation
MH  - RNA, Messenger/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - T-Lymphocytes/physiology
MH  - Transfection
EDAT- 2002/01/31 10:00
MHDA- 2002/02/15 10:01
CRDT- 2002/01/31 10:00
PHST- 2002/01/31 10:00 [pubmed]
PHST- 2002/02/15 10:01 [medline]
PHST- 2002/01/31 10:00 [entrez]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 2002 Feb;43(2):546-55.