Lung dECM microcarriers for 3D proliferation of hUCMSCs and expression of growth factors

Abstract A simple 3D proliferation strategy for human umbilical cord‐derived mesenchymal stem cells (hUCMSCs) was developed based on porcine decellularized lung tissue microcarriers (DLTMs). The lung tissue was fixed with 75% ethanol, followed by decellularization and digestion with pepsin‐acetic acid solution to form DLTMs with an average diameter of 5–40 µm. The presence of key proteins, including collagen I, collagen IV, laminin, fibronectin, and elastin, was confirmed through immunofluorescence staining, while the amount of collagen and polysaccharides was quantified. After centrifugation, DLTMs and hUCMSCs suspension formed spheroids which were cultured for 14 days. Various techniques, including DAPI, FDA and H&E staining, and DNA quantification were used to assess the performance of DLTMs on proliferation of hUCMSCs. Flow cytometric immunophenotyping was employed, and the gene and protein expressions of cytokines and growth factors such as IL‐10, HGF, KGF, and Ang‐1 were evaluated. The results demonstrated that the DLTMs exhibited a porous structure and simulated the in vivo microenvironment, effectively enhancing the proliferation and immunomodulatory functions of hUCMSCs. This strategy showed a potential for integration with bioreactor systems for large‐scale production of hUCMSC‐derived paracrine products for therapeutic applications.