Recombinant human igf 1

Human DLBCL cell lines (LY1, LY8, LY3, and Val) were routinely cultured in Iscove’s modified Dulbecco’s medium (IMDM) with 10% heat-inactivated fetal bovine serum (Gibco, MD, USA). The medium contained a 1% penicillin/streptomycin mixture and 2 mM glutamine. Verteporfin (VP; a YAP inhibitor, SML0534) was obtained from Sigma (MO, USA). Recombinant human IGF-1 was obtained from PeproTech (100-11, NJ, USA). Doxorubicin (S1208), AG1024 (an IGF-1R inhibitor, S1234), and picropodophyllin (PPP; an IGF-1R inhibitor, S7668) were purchased from Selleckchem (TX, USA).

The human HCC cell line HepG2 and Hep3B cell line were purchased from ATCC (Manassas, VA, USA). Cells were routinely cultured in DMEM (Invitrogen) supplemented with 5% fetal bovine serum (Sigma) at 37°C under 5% CO2. Recombinant human IGF-1 was obtained from PeproTech (Rocky Hill, NJ). STAT5 inhibitor was purchased from Calbiochem (San Diego, Calif., USA).

The solutions were prepared 1 to 3 days before electrospinning. For each scaffold, a polyethylene glycol (PEG) (35 kDa, Sigma-Aldrich, Berlin, Germany #81310) solution was prepared by adding 1.5 g of PEG and 3.5 g of chloroform (Sigma–Aldrich, Germany, #132950). The DP solution was produced by adding 0.6 g of DP powder, 3.52 g of chloroform, and 0.88 g of 1,1,1,3,3,3-Hexafluoro-2-propanol (HFP, Sigma-Aldrich, Germany, #105228) into a glass with a screw cap. For the incorporation of recombinant human IGF-1 (PeproTech, Boston, MA, USA, #100-11-100UG), a total amount of 4 µg of IGF-1 dissolved in 400 µL of phosphate-buffered solution (PBS, BioConcept, Allschwil, Switzerland, #3-05F39-I) containing RSA (10 µg/mL IGF-1 and 0.25% RSA in PBS) was added drop-wise to the DP solution, while stirring for five minutes on the magnetic stirrer at 500 rpm. The solution was shortly mixed on the vortex and further emulsified in an ultrasonic bath for 15 min. The emulsion was filled in a 5-mL glass syringe (Huberlab, Aesch, Switzerland,# 3.7102.33) and used immediately for electrospinning. As this protocol for water-in-oil emulsion with IGF-1 was the same as used previously for PDGF-BB incorporation, homogenous distribution of water droplets containing IGF-1 within the DP fibers can be assumed [23 (link)].