Figure 4.

Prospective isolation of mTEC and cTEC PolyKRT

(A) Workflow of thymic tissue digestion, stromal cell enrichment, and fluorescence-activated cell sorting (FACS) for cortical and medullary epithelial populations. Image created with Biorender.com. Representative FACS plots of dissociated and enriched thymic cells from human postnatal thymi (n = 13). cTECs and mTECs were gated for BCAM expression. Four TEC populations were sorted.

(B) Rhodamine-B staining of sorted cTEC and mTEC populations after two passages in culture demonstrating high clonogenic potential of BCAM^pos cells that gave rise to colonies of variable sizes that stained either strongly or dimly with rhodamine-B (n = 4, donor-derived cultures).

(C) Volcano plot analysis of freshly sorted clonogenic versus non-clonogenic thymic epithelial cells. All genes present on the StemCell NanoString nCounter Panel-Plus have been plotted. Each dot represents one gene. A p value of 0.05 and a fold change of 2 are indicated by gray, highlighting the most significantly upregulated (red) and downregulated (blue) genes (n = 3, thymic samples).

(D) Single-gene expression profiles of adhesion molecules and surface markers expressed by BCAM^pos and BCAM^neg TECs via nCounter NanoString Technologies (n= 3, thymic samples).