Fetal bovine serum growth medium

MSCs derived from normal mouse bone marrow were purchased from Cyagen Biosciences, Inc. (Guangzhou, China), and pulmonary microvascular endothelial cells (PMVECs) were purchased from Shanghai Zhen Biotechnology, Inc. (Shanghai, China). The supplier identified cells according to cell surface phenotypes or multipotency routinely. MSCs/PMVECs were cultured in DMEM/F12 or DMEM containing 10% fetal bovine serum growth medium (Wisent, China) and humidified 5% CO₂ incubator at 37°C. We changed the culture media every other day, and passage 3-7 cells were used for the experiment.

MSCs derived from bone marrow of female C57BL/6 mouse were purchased from Cyagen Bioscience, Inc. (Guangzhou, China) and general identified according to surface phenotypes and multipotency by supplier. DMEM/F12 containing 10% fetal bovine serum growth medium (Wisent, Nanjing, China) and humidified 5% CO2 incubator at 37 °C were suitable for MSC cultivate. And culture media were changed every 2–3 days. MSCs of passages 3–7 from the same batch were finally used for all experiment.
MSCs were cultured and divided into the normoxia culture group (20% O2) and hypoxia culture group (1% O2) through inflating different nitrogen. Each group received different treatments for 0 to 24 h. Before some experiments, MSC were treated with HIF-1α inhibitor KC7F2 (20 μM, Selleck, USA). For Transwell migration assay, SDF-1 (100 ng/ml, Millipore, MA) were used.