GSM5582082: human_BMSC_2D_rep3; Homo sapiens; RNA-Seq
Human bone marrow sample was obtained from the iliac crests of donors upon written consent, diluted 1:1 with Ca2+/Mg2+ HBSS, and digested with 3 mg/ml collagenase type I (Worthington Biochemical), 4 mg/ml Dispase II (Sigma) and 100 U/ml DNase I (Sigma) at 37°C for 15 min. Mononuclear bone marrow cells were isolated by Ficoll-Paque PREMIUM density gradient media (GE Healthcare, 1.073 g/ml) and cultured in low-glucose Dulbecco's modified Eagle medium (Corning, 10-014-CV) supplemented with 10% FBS (Gibco 10270-106, Lot #: 42F6480K) and 1% penicillin/streptomycin (Thermo). BMSCs were cultured in a 37°C, 5% O2 and 5% CO2 incubator. 80,000 2D expaned and 3D cultured BMSC in GelMA for 7 days were dissociated, total RNA was extracted, digested with DNase I and purified using RNAclean Kit. RNA-seq libraries were constructed by a modified Smart-seq2 method with UMI barcodes (Nature 533:487, 2016).