Single cell analysis of murine lung epithelial cells cultured in air-liquid inteface cultures

Typ 2 pneumocyte proliferation and differentiation is required for pulmonary homeostasis and tissue regeneration. A disturbed crosstalk between fibroblasts and pneumocytes contributes to the loss of lung structure in chronic lung diseases, such as COPD and IPF. Therefore, it is important to understand the role of fibroblasts in pulmonary epithelial regeneration. Here, we show that primary fibroblasts drive the differentiation of murine pneumocytes in air-liquid interface cultures. Co-cultures of primary pneumocytes with fibroblasts resulted in a significantly increased trans-epithelial resistance and increased expression of Type 2 and Type 1 markers. Single cell sequencing revealed that fibroblasts specifically activate signaling pathway (e.g. IGF pathways) in epithelial cells. Our co-culture model is useful for studying cellular mechanisms that mediate the fibroblasts-driven differentiation of pneumocytes.