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Three-dimensional collagen matrices as cell culture substrata affect the generation of alloreactive cytotoxic T
1Department of Microbiology, Kitasato University School of Medicine, Kanagawa, Japan.
Primary one-way mixed lymphocyte cultures (MLC) of C3H/He responder and DBA/2 stimulator were performed in three-dimensional (3-D) collagen matrices and the generation of alloreactive cytotoxic T cell (CTL) responses was compared to those in MLC which were done on usual plastic surfaces or on collagen-coated plastic surfaces. MLC in the 3-D collagen matrices were found to generate strong CTL responses. Flow cytometric analysis of Lyt-2 and L3T4 antigen expressions on the effector cells showed that the Lyt-2/L3T4 ratios were substantially higher in the 3-D collagen matrices, and that a larger proportion of the cells in the 3-D collagen matrices were Lyt-2+ lymphoblasts. These results indicate that the milieu of the 3-D collagen matrices favors the proliferation of Lyt-2+ lymphocytes, and suggests that cell-to-matrix interactions in 3-D collagen matrices may play a regulatory role in the maturation process of alloreactive CTLs.
Primary one-way mixed lymphocyte cultures (MLC) of C3H/He responder and DBA/2 stimulator were performed in three-dimensional (3-D) collagen matrices and the generation of alloreactive cytotoxic T cell (CTL) responses was compared to those in MLC which were done on usual plastic surfaces or on collagen-coated plastic surfaces. MLC in the 3-D collagen matrices were found to generate strong CTL responses. Flow cytometric analysis of Lyt-2 and L3T4 antigen expressions on the effector cells showed that the Lyt-2/L3T4 ratios were substantially higher in the 3-D collagen matrices, and that a larger proportion of the cells in the 3-D collagen matrices were Lyt-2+ lymphoblasts. These results indicate that the milieu of the 3-D collagen matrices favors the proliferation of Lyt-2+ lymphocytes, and suggests that cell-to-matrix interactions in 3-D collagen matrices may play a regulatory role in the maturation process of alloreactive CTLs.