3D hydrogel culture system for generation of fibrosis-associated macrophages

We identified a subpopulation of macrophages that are specifically enriched in interstitial lung disease. To better understand the functional relevance of this population with regards to disease pathogenesis by manipulating them in vitro, we attempted to induce this state in cell culture by prepurposing a published 3D hydrogel culture system originally designed to generate hematopoietic progenitor cells from PBMCs. Whole blood from three healthy donors was collected. PBMCs were extracted and put into the 3D hydrogel culture system described by Xu et al. (doi:10.1007/s13238-021-00900-4), cultured for 8 days, and then extracted from the hydrogel for scRNA-seq. Donors were pooled for scRNA-seq to be later computational separated by genetic demultiplexing.