Fig. 4.

Applicability of the tissue-engineered model to human FRCs: Characterization of PLN FRCs from healthy human donors in LNs and of sorted FRCs cultured in 3D collagen scaffolds. (a) Representative maximum projection image of confocal z-stacks of human PLNs from a healthy donor. FRCs are identified by the FRC marker gp38 (green). Scalebar = 50 µm. (b) Reticular pore size (Feret diameter) of human FRC networks in PLNs compared to PLNs from 12-week old NOR mice (n = 3 human PLN donors; n = 3 NOR PLN donors). For each PLN, n = 30 pores were quantified (c) Gating strategy for FACS sorting of gp38⁺ CD31⁻ LN cells after depletion of CD45⁺ cells isolated from enzyme-based digestion of PLNs as previously reported²⁵. (d) Representative maximum projection images of confocal z-stacks taken 1–5 days after seeding 1 × 10⁵ human PLN FRCs in collagen sponges. FRCs are identified by gp38 (green), and F-actin (red); nuclei are counterstained with DAPI (blue). Scalebars = 100 µm. *p < 0.05