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Re: A deleted message

Wednesday, 03/16/2022 3:47:13 PM

Wednesday, March 16, 2022 3:47:13 PM

Post# of 693125
Flip a quick question.

There has been a lot of talk about Microden but very little about EDEN which is the large scale producer of DC's.

Here is some material I found regarding EDEN and MicroDen.

I would assume NWBO also owns the rights to EDEN.

If we are all aware of this please ignore.

From Article:

Correspondence: Shashi Murthy (s.murthy@northeastern.edu) Background
Dendritic cells (DCs) are effective vehicles for personalized therapies and there is substantial interest in leveraging their capabilities for improving patient outcomes [1,2]. However, the process of generating these DCs, typically from monocytes (MOs), in an efficient and reproducible process has proved challenging. We have developed automated closed systems, EDEN (large scale) and MicroDEN (small scale), for generating clinically relevant numbers of DCs from precursor MOs which can be sterile-transferred for downstream applications. These systems overcome many of the constraints of manual processing and are designed for the research environment as well as clinical manufacturing.

Methods

Enriched MOs were seeded into EDEN at a density of 206,000 MOs/cm2 using RPMI 1640 medium supplemented with 10% HI-FBS and 500 U/mL IL-4 and GM-CSF (R&D). Medium was continuously perfused and immature DCs were harvested on Day 6.Enriched MOs were seeded into MicroDEN at densities of 200k, 400k, and 600k MOs/cm2 using CellGenix DC Medium supplemented with 350 U/mL IL-4 and GM-CSF (CellGenix). Medium was continuously perfused and DCs were harvested on Day 6.

Results

Flow cytometry indicated that immature DCs (iDCs) from EDEN exhibited standard DC-SIGN (CD209), CD14, and CD80/83/86 (Figure 2). Antibody expression is comparable to prior results collected in MicroDEN and well plate controls. Approximately 25 million viable iDCs were harvested from EDEN for an iDC yield of 32% with the relatively low seeding density. These results indicate that EDEN can be used to generate clinically relevant numbers of iDCs in a single closed system. Flow cytometry indicated that MicroDEN generated phenotypically similar iDCs at each seeding density with an iDC yield of ~30% (Figure 3), similar to well plate controls. Allogeneic functional assays indicated that MicroDEN iDCs exhibited improved proliferative activity than well plate iDCs and generally exhibited better activity at lower seeding densities (Figure 4).

Conclusions

DCs generated in both EDEN and MicroDEN are phenotypically comparable and exhibit increased proliferative activity to standard manual culture. Upto 26 million DCs can be generated by MicroDEN at the highest seeding density whereas the output of EDEN is 26 million DCs at the lowest seeding density.
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