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Human retinal stem cells secreting CNTF (National Eye Institute)

NTC200 is an established human retinal pigment epithelium cell line. These cells are transfected with ciliary neurotrophic factor (CNTF), which is then produced by the cells and secreted to the extra-cellular medium. Two stably transfected sub-lines exist: a high-CNTF-secreting line designated NTC-201-6A and a low-CNTF-secreting line designated NTC-201-10.

Human retinal pigment epithelium cell line ARPE-19 (catalog no. CRL-2302; American Type Culture Collection) were transfected with the human CNTF gene. Stable cell lines were selected by exposure to geneticin, at a concentration of 1.0 mg/mL, and later maintained in 0.25 mg/mL geneticin. A high CNTF-secreting cell line (1.3±0.2 gene copies/cell) and a low CNTF-secreting cell line (0.6±0.1 gene copies/cell) were generated by varying the amount of CNTF plasmid used in the transduction. Stable cell lines were established and designated NTC-201-6A (800ng/106 cells/day in vitro) and NTC-201-10 (250ng/106 cells/day in vitro). Cell lines are maintained in DMEM–F12 supplemented with fetal bovine serum (10%) in a 5% CO2, 95% humidity, 37°C incubator.

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Homo sapiens
Human retinal stem cells secreting CNTF