3D engineered scaffold for large-scale Vigil immunotherapy production.

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.

Neurotrophic support by traumatized muscle-derived multipotent progenitor cells: Role of endothelial cells and Vascular Endothelial Growth Factor-A.

BACKGROUND: Adult mesenchymal stem cells (MSCs) have been shown to increase nerve regeneration in animal models of nerve injury. Traumatized muscle-derived multipotent progenitor cells (MPCs) share important characteristics with MSCs and are isolated from severely damaged muscle tissue following surgical debridement. Previous investigations have shown that MPCs may be induced to increase production of several neurotrophic factors, suggesting the possible utility of autologous MPCs in peripheral nerve regeneration following injury. Recent findings have also shown that components of the vascular niche, including endothelial cells (ECs) and vascular endothelial growth factor (VEGF)-A, regulate neural progenitor cells and sensory neurons. METHODS: In this study, we have investigated the neuroinductive activities of MPCs, particularly MPC-produced VEGF-A, in the context of an aligned, neuroconductive nerve guide conduit and the endothelial component of the vascular system. Embryonic dorsal root ganglia (DRG) seeded on poly-ϵ-caprolactone aligned nanofibrous scaffold (NF) constructs and on tissue culture plastic, were cocultured with induced MPCs or treated with their conditioned medium (MPC-CM). RESULTS: Increased neurite extension was observed on both NF and tissue culture plastic in the presence of MPC-CM versus cell-free control CM. The addition of CM from ECs significantly increased the neurotrophic activity of induced MPC-CM, suggesting that MPC and EC neurotrophic activity may be synergistic. Distinctly higher VEGF-A production was seen in MPCs following neurotrophic induction versus culture under normal growth conditions. Selective removal of VEGF-A from MPC-CM reduced the observed DRG neurite extension length, indicating VEGF-A involvement in neurotrophic activity of the CM. CONCLUSIONS: Taken together, these findings suggest the potential of MPCs to encourage nerve growth via a VEGF-A-dependent action, and the use of MPC-CM or a combination of MPC and CM from ECs for peripheral nerve repair in conjunction with NFs in a nerve guide conduit. Due to the ease of use, application of bioactive agents derived from cultured cells to enhance neurotrophic support presents a promising line of research into peripheral nerve repair.