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. 2024 Jul 5;14(1):15556.
doi: 10.1038/s41598-024-65993-3.

3D engineered scaffold for large-scale Vigil immunotherapy production

Fabienne Kerneis  1 , Ernest Bognar  1 , Laura Stanbery  1 , Seongjun Moon  2 , Do Hoon Kim  2 , Yuxuan Deng  2 , Elliot Hughes  2 , Tae-Hwa Chun  2   3 , Darron Tharp  1 , Heidi Zupanc  1 , Chris Jay  1 , Adam Walter  1   4 , John Nemunaitis  5 , Joerg Lahann  2
Affiliations

3D engineered scaffold for large-scale Vigil immunotherapy production

Fabienne Kerneis et al. Sci Rep. .

Abstract

Previously, we reported successful cellular expansion of a murine colorectal carcinoma cell line (CT-26) using a three-dimensional (3D) engineered extracellular matrix (EECM) fibrillar scaffold structure. CCL-247 were grown over a limited time period of 8 days on 3D EECM or tissue culture polystyrene (TCPS). Cells were then assayed for growth, electroporation efficiency and Vigil manufacturing release criteria. Using EECM scaffolds, we report an expansion of CCL-247 (HCT116), a colorectal carcinoma cell line, from a starting concentration of 2.45 × 105 cells to 1.9 × 106 cells per scaffold. Following expansion, 3D EECM-derived cells were assessed based on clinical release criteria of the Vigil manufacturing process utilized for Phase IIb trial operation with the FDA. 3D EECM-derived cells passed all Vigil manufacturing release criteria including cytokine expression. Here, we demonstrate successful Vigil product manufacture achieving the specifications necessary for the clinical trial product release of Vigil treatment. Our results confirm that 3D EECM can be utilized for the expansion of human cancer cell CCL-247, justifying further clinical development involving human tissue sample manufacturing including core needle biopsy and minimal ascites samples.

Keywords: Autologous tumor cell therapy; Cell expansion; Immunotherapy; Ovarian cancer; Vigil.

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Conflict of interest statement

This collaborative study has been supported by Gradalis. The University of Michigan has filed a patent application related to aspects of this work. JN is a board member and employee of Gradalis Inc. FK, EB, LS, HZ, CJ, and AW are employees of Gradalis, Inc. All other authors do not hold any competing interest.

Figures

Figure 1
Figure 1
Proliferation curves for CCL-247 (A) and CT-26 (B) cells. Cells were grown on 3D EECM or TCPS for 8 days. The cell proliferation curve was quantified using the TOX8 assay daily.
Figure 2
Figure 2
Transfection Efficiency of CCL-247 cells. Cells were transfected with GFP plasmid. The percentages of GFP-expressing cells were determined by flow cytometry. (A) Electroporated cells cultured on 3D EECM without GFP plasmid, (B) Electroporated cells cultured on 3D EECM with GFP plasmid, (C) Electroporated cells cultured on TCPS with GFP plasmid.
Figure 3
Figure 3
(A) Cell growth on scaffold matrix under 10× phase contrast microscopy at Day 2 and (B) Day 6.
Figure 4
Figure 4
Vigil manufacturing process.
Figure 5
Figure 5
GM-CSF (A) and TGF-β1 (B) Cytokine Expression level pre-treatment (Pre TFX) versus post irradiation (Post IRR). *p = 0.0001, **p < 0.000001.
See this image and copyright information in PMC

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