Mechano-biomimetic hydrogel 3D cell cultivation as a strategy to improve mammalian cell protein expression.

Eukaryotic expression systems are frequently employed for the production of recombinant proteins as therapeutics as well as research tools. Among which mammalian cell protein expression approach is the most powerful one, which can express complex proteins or genetic engineered biological drugs, such as PD-1. However, the high expense, which partially derives from its low protein yielding efficiency, limited the further application of such approach in large scale production of target proteins. To address this issue, we proposed a novel technique to promote the protein production efficiency of mammal cells without using conventional genetic engineered approaches. By placing 293T cells in a hydrogel 3D cell culture platform and adjusting the stress relaxation of the matrix hydrogel, cells formed multicellular spheroids by self-organization. In particular, the multicellular spheroids have a significantly enhanced ability to transiently express multiple proteins (SHH-N, PD-1 and PDL-1). We also examined in detail the mechanism underlying this phenomenon, and found that the reorganization of cytoskeleton during spheroids formation enhances the translation process of protein by recruiting ribosomes. Overall, this finding provides a novel approach for subsequent improvement of large-scale mammalian protein expression cell systems.

Evaluation of joint effects of perfluorooctane sulfonate and wood vinegar on planarians, Dugesia japonica.

Perfluorooctane sulfonate (PFOS) is a persistent organic pollutant and can cause oxidative stress in animals. Wood vinegar (WV) is the water condensate of smoke produced during wood carbonization. It was used for antibacterial application, pest control, and antioxidant. In the study, PFOS and WV were used to treat the planarian, and then the oxidative stress induced by PFOS on the planarian (Dugesia japonica) and the protective effects of WV on lipid peroxidation, related antioxidant enzyme activity, and mRNA expression in the planarian were studied. PFOS caused an increase in malondialdehyde (MDA) contents, a decrease in superoxide dismutase (SOD) and catalase (CAT) activities, and a change in glutathione peroxidase (GPx), glutathione S-transferase (GST), glutathione reductase (GR) activities. The mRNA levels of glutathione peroxidase gene (gpx), glutathione S-transferase enzyme gene (gst), and glutathione reductase gene (gr) are upregulated or downregulated to varying degrees. The WV and co-treatment planarians reduced MDA levels, increased the activities of oxidative stress biomarker enzymes, and restored gene expression levels. Our results show that low concentration of WV has protective effects on the oxidative damage caused by PFOS in the planarian.