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Cahil McGovern, Sandy Sherman, Crystal Cortellessa, Suzanna Borges, Melissa Stiles, Karen Ahola, Alice Lee, Konrad Kauper, Bruce Bouchard, Weng Tao; Evaluation of a Candidate Cell Line Employed to Deliver an Antiangiogenic Factor Using Encapsulated Cell Technology. Invest. Ophthalmol. Vis. Sci. 2012;53(14):465.
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To investigate the cell stability and in vitro performance of Encapsulated Cell Technology (ECT) devices using a cell line to deliver an antiangiogenic factor.
The cell line secreting the antiangiogenic factor was constructed using NTC-200 cells. The candidate cell line was cultured for 40 passages. At predetermined passages, cells were seeded at a defined density in 24 well plates and allowed to attach. Cells were washed twice with a balanced salt solution and incubated with 1 ml of Endo-SFM for 2 hours. The pulsate was then analyzed for antiangiogenic factor release. Once cell stability had been established out to 40 passages, the cell line was encapsulated in a hollow fiber membrane. Each device was manufactured using standard protocols and held at 37°C in closed packages containing 37 mls of Endo-SFM. ECT devices were pulsed for factor production in 1 ml of Endo-SFM for 24 hours at days 2, 7, and 14 post manufacture. The devices were subsequently analyzed for metabolic activity and then subjected to either total DNA or histological analysis. Unencapsulated cells and device performance were quantified by Elisa. Device metabolic activity was determined using the CCK-8 assay (Dojindo). Total DNA was determined using the Hoefer DyNA Quant 200 fluorometer (Pharmacia). Histological examination of the devices was performed using standard hematoxylin and eosin staining techniques.
Cell stability: The candidate cell line released the desired factor for 40 passages which is favorable for manufacturing purposes. During this time, the cells maintained a typical and consistent morphology as well as exhibited a consistent doubling rate. Device stability: The ECT devices released the desired factors and maintained viable cells during the evaluation period. Total DNA analysis of devices showed a consistent number of cells was maintained between 1 and 2 weeks and histological analysis of device sections showed a high density of healthy cells distributed throughout the device.
The data suggested that the Encapsulated Cell Technology platform was able to achieve sustained delivery of antiangiogenic factors under in vitro conditions. This technology can deliver other factors for a broad range of indications where long-term treatment is required.
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