Rvegf

Rat BMSCs were isolated from the femurs and tibias of male Wistar rats (90–100 g) as described earlier with modification [24 (link)]. Rats were killed by intravenous injection of ketamine/xylazine, and all work was performed according to the Institutional Animal Care and Use Committee guidelines. This experimental protocol was reviewed and approved by the Medical Ethics Committee of Shandong University School of Medicine. Briefly, the cells were seeded in Dulbecco's modified Eagle's medium low glucose (Gibco, USA) supplemented with 15% fetal bovine serum and fibroblast growth factor 2 (FGF-2) (Beifuji, China), 10 ng/mL, at 37°C in humidified air with 5% CO₂. Rat BMSCs were phenotypically characterized as described [25 (link)]. Cells were transfected with pCMV6 empty vector/pCMV6-CD163 in DMEM with FGF-2 (10 ng/mL), rVEGF (10 ng/mL) (Sigma, St. Louis, MO), and 2% serum and incubated at 37°C for 24 h. Then, the medium is replaced by basal DMEM with FGF-2 (10 ng/mL) and rVEGF (10 ng/mL) incubated at 37°C for another 24 h.

The A platensis was purchased from Martin Bauer GmbH (Vestenbergsgreuth, Germany). The water extract of both A platensis and phycocyanobilin was prepared as has been previously described elsewhere.4 The cell culture media and non‐essential amino acids (NEAAs) were obtained from Sigma‐Aldrich, and the other cell culture components were from Biosera (Nuaille, France). The serine/threonine phosphatase and protease inhibitor cocktails were purchased from either Sigma‐Aldrich or Serva. The Geltrex™ LDEV‐Free Reduced Growth Factor Basement Membrane Matrix was purchased from Thermo Fisher Scientific. The recombinant growth factors and inhibitors were procured as follows: rVEGF, rEGF (epidermal growth factor), rAREG (amphiregulin, autocrine mitogen related to EGF), rHGF/SF (hepatocyte growth factor/scatter factor), PD 0325901 (all from Sigma‐Aldrich), erlotinib (Cell Signaling Technology), vatalanib and axitinib (Selleck Chemicals) and bevacizumab (LGM Pharma). Unless otherwise specified, all other common chemicals were from Sigma‐Aldrich.