Human msc growth medium

Human umbilical cord MSCs (hucMSCs) were purchased from Cyagen (Suzhou, China), and human dermal fibroblasts (HDFs) from Jennio (Guangzhou, China). hucMSCs were cultured in human MSC growth medium (Cyagen) and passaged when the confluency reaches approximately 80–90%. At passage 6, the culture medium was replaced with complete MesenCult™-ACF Plus Medium (STEMCELL Technologies, Hong Kong, China), which is a serum- and animal component-free culture medium, and collected for the isolation of exosomes at 90% cell confluency. HDFs were cultured and expanded in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum, penicillin (100 U/mL) and streptomycin (100 mg/mL), and used for the experiments over passages 5–9. All these cells were cultured in a humidified incubator under a 5% CO₂ atmosphere at 37°C.

Human adult BM-MSCs were obtained from Cyagen Biosciences (Jiangsu, China; passage 2, Cat. No. HUXMA-01001) and cultured in human MSC growth medium (Cyagen Biosciences) supplied with 10% fetal bovine serum (FBS), 1% penicillin–streptomycin, and 1% glutamine. Human bronchus epithelial cells BEAS-2B were purchased from ATCC (Rockville, MD) and cultured in DMEM/F12 with 10% FBS and 1% penicillin–streptomycin. Human embryonic kidney cells 293T (ATCC) were cultured in Dulbecco's modified Eagle medium (DMEM) supplied with 10% FBS and 1% penicillin–streptomycin. All cells were cultured in a humidified chamber under 5% CO₂ at 37°C.