Two high-yield complementary methods to sort cell populations by their 2D or 3D migration speed

Abstract

The potential to migrate is one of the most fundamental functions for various epithelial, mesenchymal and immune cells. Image analysis of motile cell populations, both primary and cultured, typically reveals an intercellular variability in migration speeds. However, cell migration chromatography, the sorting of large populations of cells based on their migratory characteristics, cannot be easily performed. The lack of such methods has hindered our understanding of the direct correlation between the capacity to migrate and other cellular properties. Here, we report two novel, easily implementable and readily scalable methods to sort millions of live migratory cancer and immune cells based on their spontaneous migration in 2D and 3D microenvironments respectively. Correlative downstream transcriptomic, molecular and functional tests reveal marked differences between the fast and slow subpopulations in patient-derived cancer cells. We further employ our method to reveal that sorting the most migratory cytotoxic T lymphocytes yields a pool of cells with enhanced cytotoxicity against cancer cells. This phenotypic assay opens new avenues for the precise characterization of the mechanisms underlying hitherto unexplained heterogeneities in migratory phenotypes within a cell population, and for the targeted enrichment of the most potent migratory leukocytes in immunotherapies. [Media: see text] [Media: see text] [Media: see text]

BMP-2-immobilized PCL 3D printing scaffold with a leaf-stacked structure as a physically and biologically activated bone graft

“…This finding was consistent with the study of Lecanda et al [57], who reported that BMP-2 affected the differentiation of human MSCs in osteogenic medium, but was not significantly involved in proliferation. The rapid cell proliferation on the smooth surface than on LSS surface after 1 week of cell culture can be attributed to the change of PCL surface into an interactive surface by the adsorption of cell adhesive proteins in the cell culture medium and accelerated migration and proliferation to cell-free surface via interaction with surrounding cells [58][59][60][61]. To further evaluate the topographical effects of LSS on cell adhesion, the expression of focal adhesion markers of hPDCs adhering to the LSS surfaces (3D-PLSS and 3D-PLSS-BMP) was compared with those adhering to smooth surface groups (3D-PS and 3D-PS-BMP) (figures 5 and S2).…”