NorthWest Biotherapeutics Inc (NWBO)

[Smokey21]

Thanks for posting this MicroDEN abstract Sukus. The full paper is a very interesting read.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5946316/

Some interesting tidbits:

..."the current approach to DC generation poses an unusually large burden, most significantly in terms of cost, but also in terms of time required to perform comprehensive studies and trials and increased potential for error (e.g., contamination, patient cell misidentification, etc.)."

"The static culture is currently performed in either well plates or cell culture flasks (T-flasks) for both research and therapeutic purposes and requires several manually performed steps where the culture vessel is (a) open to surrounding air within an aseptic environment (e.g., laminar flow hood) and (b) transferred from an aseptic environment to an incubator. These aforementioned steps introduce potential for contamination when the incubator and laminar flow hood are not within the same aseptic environment (i.e., advanced research labs) and rely on the manual manipulation of the culture flask/cells by lab personnel."


"While the number of DCs generated in static culture varies widely, obtaining therapeutically relevant numbers of DCs requires tens of well plates or flasks even with the best DC yield. Furthermore, these protocols require cytokine replenishment every 2–3 days of culture;28, 29 this means removing the culture flask from the incubator, manually adding DC differentiation medium to each well or flask, and returning the flask to the incubator. While possible, this process becomes overly laborious when cells from multiple patients are being cultured simultaneously, placing a strain on laboratory resources and personnel time."

Upon identifying the aforementioned deficiencies in the current DC generation process, we developed an automated fluidic system that allows for differentiation of monocytes into iDCs utilizing continuous perfusion of differentiation media within an enclosed aseptic environment called MicroDEN. Manual steps associated with current ex vivo DC generation are vastly reduced with MicroDEN and an aseptic environment is ensured by the use of an enclosed DC generation cartridge and tubing network that supplies fresh cytokines and media to the cells while concurrently removing spent media from the cartridge, all of which is an advancement from current static culture techniques. Furthermore, the automated perfusion system requires no user intervention after setup and can be left to run until harvest. Benchmark phenotyping was performed on the generated iDCs along with allogeneic PBMC and syngeneic antigen functional assays. MicroDEN generated iDCs were phenotypically similar to well plate generated iDCs and there were no salient differences between MicroDEN and well plate generated iDCs in functional assays developed to study DC-dependent T-cell induction.

This is Huge!!!