Supplementary Figure 5: Enhanced positive selection in ATOs requires 3D structure and optimal cell line and culture medium. | Nature Methods

Supplementary Figure 5: Enhanced positive selection in ATOs requires 3D structure and optimal cell line and culture medium.

From: Generation of mature T cells from human hematopoietic stem and progenitor cells in artificial thymic organoids

Supplementary Figure 5

Enhanced T cell positive selection and maturation in ATOs (defined as MS5-hDLL1 in 3D culture with RB27) compared with monolayer co-cultures. Week 6 monolayer cultures (left) were compared with 3D organoid cultures (right) and included crossover comparisons with either MS5-DL1 or OP9-DL1 cells and RB27 medium or OP9-DL1 standard medium as indicated. Standard medium for OP9-DL1 co-cultures was MEMα/20%FCS with IL-7, FLT3L, and ascorbic acid, and standard medium for ATOs was RB27 with IL-7, FLT3L, and ascorbic acid, as described in Methods. Monolayer or 3D cultures using the parental MS-5 cell line (not transduced with DLL1) are also shown as a negative control. All plots are gated on CD14-CD56- cells or (where indicated) CD3+TCRαβ+ subgates.

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