Abstract
Human alveolar type 2 (AT2) cells play a crucial role in maintaining lung homeostasis and facilitating repair. Researchers are investigating AT2 cells in ex vivo platforms due to their essential roles in lung physiology, disease modeling, and drug testing. Herein, we present a comprehensive workflow for the isolation and culture of primary human AT2 cells, including an efficient enzyme-based lung tissue digestion protocol and automated purification of AT2 cells from both healthy and diseased lungs. By targeting the HTII-280 surface protein on AT2 cells, we employed an automated magnetic-activated cell sorting (MACS) separation strategy using the autoMACS® Pro Separator. AT2 cells from healthy and diseased lungs were grown in 3D culture and formed alveolospheres, elucidating that the isolation strategy did not affect cell functionality. This approach provides high AT2 cell yield and purity, offering scalability to generate sufficient quantities of cells for downstream applications, such as disease modeling and cell-based therapeutics.