- Authors: Chae YJ. et al.
- Year: 2019
- Journal: Transl Oncol 12 1264-1270
- Applications: in vitro / DNA / FectoPRO
- Cell type: HEK-293F
These two vectors were purified using a DNA Maxi-prep kit from Qiagen and transfected into HEK293F cells at a 1:2 DNA ratio (heavy chain: light chain) using FectoPRO transfection reagent (Polyplus transfection) in accordance with the manufacturer's instructions. After 10 days of antibody production in ‘FreeStyle 293’ expression medium, supernatant containing the antibodies was harvested and purified using open-column chromatography with protein A resin
Immune checkpoint inhibitors (ICIs) have become an effective therapeutic option for colorectal cancer and studies on these drugs have therefore increased greatly. Efficacy assessments of ICIs in preclinical orthotopic colorectal cancer using MRI have not been reported however due to the difficulties in conducting colorectal imaging. The purpose of this present study was to investigate the feasibility of using magnetic resonance colonography (MRC) to evaluate the efficacy of an ICI, an anti-PD-L1 antibody, in an orthotopic colorectal cancer mouse model. The mouse model was generated by the engraftment of colorectal cancer cells into the submucosal layer of the colon. Anti-cancer efficacy was assessed by tumor volume and metastatic tumor number analyses, and these values were significantly lower in the PD-L1 antibody-treated group compared to the controls. Histological analyses using H&E and Ki-67 immunohistochemical staining confirmed a highly efficacious tumor growth inhibition and enhanced infiltration by CD4+ and CD8+ lymphocytes in the PD-L1 antibody-treated group. We conclude that MRC has the potential to be used for ICI efficacy assessments against orthotopic colorectal cancer mouse model.