title: NextSeq 500 sequencing; RNAseq of human bone marrow mesenchymal stem cells in co-culture with endothelial cells
design: RNAseq of human bone marrow mesenchymal stem cells in co-culture with endothelial cells
library_construction_protocol: MSCs monocultures or MSC / EC cocultures were encapsulated in 1.7% TG-PEG hydrogels (470Pa) to generate three-dimensional cultures. 3D cultures were maintained under standard culture conditions for 1 week in MEM Alpha containing 10%FBS, Pen/Strep and 50ng/mL FGF-2. Hydrogels were digested by collagenase treatment and MSCs were isolated by FACS sorting based on the absence of CD31 marker Expression.  Endothelial cells for the sequencing experiment were human umbilical vein endothelial cells (PromoCell). These cells are also primary cells. RNA was isolated using RNeasy Micro Kit (Qiagen) RNA Quality was assessed using Agilent TapeStation 4200 and RNA quantity was determined using RiboGreen (Life Technologies) and measured on a Infinite M1000 Pro plate Reader (Tecan). 200ng RNA was used in a TrueSeq Stranded mRNA Library Prep Kit (Illumina) to generate a RNA-seq library.
attributes.tag: Experimental Factor: growth condition
attributes.value: monoculture