GSM6857898: BMP2 rep 3; Homo sapiens; RNA-Seq
Briefly, Gelfoam gelatin sponges (Pfizer, Cat.# 0009-0323-01) were sectioned into 48 pieces, washed with ethanol 70% (Scharlab, Cat.# ET0003005P) and rehydrated in sterile PBS (Gibco, Cat.# 14040). hBM-MSCs were diluted in culture media at 1x106 cells/ml and 100microl (1x105 cells) were carefully inoculated in each scaffold using a 1mL syringe (Braun, Cat.# 9161406V) with 25G needle (Braun, Cat.# 9186166).  Cell-seeded scaffolds were transferred to polystyrene ultra-low attachment 24 well plates (Corning, Cat.# 3473) and maintained in cell culture conditions for 3-5 hours. Then, culture media was added and scaffolds were cultured for 3 to 7 days. 5 microL of acetic acid (Fluka, Cat.# 27225) 50 mM reconstituted rhBMP-2 (Noricum, Cat.# rhBMP-2) (concentration at 5 microg/microL) were added. Then, 30 microL of 2% CaCl2 reconstituted thrombin from human plasma (Merck, Cat.# T8885) and 30 microL of water reconstituted fibrinogen from human plasma (Merck, Cat.# F3879) were incorporated. RNA libraries were prepared for sequencing using standard Illumina protocols