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W. Tente, C.G. Thanos, K. Kauper, S. Sherman, W. Bell, R. Provencal, W. Tao; The Effect of Ambient Storage Conditions on the Recombinant Protein Secretion Rates and Cell Viability of Polymer Encapsulated Human Retinal Pigmented Epithelial Cells Genetically Engineered to Secrete Ciliary Neurotrophic Factor . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4435.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: The Neurotech NT-501 cell-encapsulated device for intraocular delivery of CNTF to the posterior cavity of the eye has been evaluated in vitro under a variety of temperature and holding conditions. This study examines devices containing either NTC-201-6A cells (high producer) or NTC-201-10 cells (low producer) over a wide range of ambient temperature conditions and holding periods for the purpose of evaluating transportation feasibility to the clinical site. Methods: NT-501 devices were manufactured with an average length of 1.1 cm, sterilized, and loaded with either of the two cell lines. Following a two-week hold period at 37°C in a closed container with 40 mL endothelial serum-free media (ENDO-SFM), devices were incubated in environmental chambers at temperature conditions ranging from 3°C to 48.4°C including variable temperature cycles. Several groups were cycled between the temperature extremes and then allowed to remain at room temperature for 1 week. At the end of the incubation period, CNTF production was analyzed by incubating the device in fresh ENDO-SFM for 24 hours followed by detection with an ELISA. Cell density was assessed using image analysis on plastic sections generated after the 24-hour pulse. Results: CNTF levels for both groups were generally unaffected by temperature variations, although the 6A group showed declining CNTF secretion and cell density above 44°C. Cell density in both groups was consistent with CNTF output and devices showed favorable morphology throughout all conditions, excluding temperatures above 44°C. Device output and histology remained optimal even following the 1-week room temperature hold after the most extreme simulated shipping cycle. Conclusions: The NT-501 device is extremely robust and able to withstand extremely variable ambient conditions, including cycles between temperature extremes. A shipping package has been designed in parallel with these studies to ensure the delivery of active devices to the clinic.
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