Fig. 1
From: High-throughput isolation of normal and pulmonary fibrosis human alveolar epithelial cells
Schematic overview of the lung digestion and cell isolation process, with alveolosphere culture and downstream analyses. (A) AT2 cells were extracted from lung tissue using enzymatic and mechanical dissociation, followed by mincing and filtering steps to achieve single cells for counting (the pre-MACS sample). (B) Using either a 1-step (HTII-280 positive enrichment) protocol or a 2-step (CD45 depletion, followed by HTII-280 positive enrichment) protocol, AT2 cells were labeled for MACS separation based on surface antigen HTII-280 and isolated using the autoMACS Pro Separator. (C) Following HTII-280 enrichment, healthy and IPF-diseased AT2 cells were counted, mixed with Matrigel, and seeded as droplets for 3D culture. After one month of growth, mature alveolospheres were collected for downstream analyses. The alveolospheres were either prepared for immunostaining and imaging or dissociated into single cells for real-time quantitative PCR (qPCR) to assess gene expression.
