Generation and characterization of immortalized rat retinal microglial cell lines.
Retinal microglia play an important role as resident immunocompetent and phagocytic cells in the event of injury and disease. Retinal microglia and microglia precursor transplantation show a rescue effect in ischemic retina and retinal degeneration. However, studies of retinal microglia have been hampered by the difficulty of obtaining sufficient numbers of microglia. One way to circumvent this difficulty is to establish permanent retinal microglia cell lines. In the present study, we report the generation of immortalized retinal microglia, T-MG cells, from postnatal day 3 rat retinal tissue using a lentiviral vector encoding SV40 large T antigen. The T-MG cells exhibited cell-type-specific antigens for monocyte/macrophage lineage cells, including CD11b (OX42), ED1 (OX6), and Iba1, and actively phagocytosed latex beads. In addition to primary retinal microglia, T-MG cells also have the ability to recruit into chemokines. Treatment of T-MG cells with lipopolysaccharide (LPS) led to increased levels of tumor necrosis factor-α, interleukin-1β, and inducible nitric oxide synthase. Genome-wide microarray analysis showed a less than 1% difference in the genes between the T-MG cells and the control primary retinal microglia. The T-MG cells exhibited properties similar to those of the primary retinal microglia and should have considerable utility as an in vitro model for the study of retinal microglia in health and as a curative therapy and an in vivo model for the study of retinal microglia in disease.
Department of Ophthalmology, West China Hospital of Sichuan University, Chengdu, Sichuan, People's Republic of China; Department of Ophthalmology, Eye and ENT Hospital of Fudan University, Shanghai, People's Republic of China; Shanghai Key Laboratory of Visual Impairment and Restoration, Fudan University, Shanghai, People's Republic of China.
Xiao-shuang
XS
Jiang
Ying-qin
YQ
Ni
Tian-jin
TJ
Liu
Meng
M
Zhang
Rui
R
Jiang
Ge-zhi
GZ
Xu
D000818:Animals; D000831:Animals, Newborn; D039481:CD11b Antigen / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002461:Cell Line, Transformed / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation; D018932:Chemokine CCL2 / Q000494:pharmacology; D002633:Chemotaxis / Q000502:physiology; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D053331:Ectodysplasins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology; D005822:Genetic Vectors; D008840:Microfilament Proteins / Q000378:metabolism; D017628:Microglia / Q000502:physiology*; D009569:Nitric Oxide / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000166:cytology*; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism; D014161:Transduction, Genetic
C100745:Aif1 protein, rat; D039481:CD11b Antigen; D002135:Calcium-Binding Proteins; D018932:Chemokine CCL2; D016207:Cytokines; D053331:Ectodysplasins; D008840:Microfilament Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D009569:Nitric Oxide; C022616:thiazolyl blue
SV40 large T-antigen; chemokines; cytokines; immortalized cell line; retinal microglia