Time course profile of umbilical cord blood cells in culture

Despite the importance of inter-cellular communication networks in regulating stem cell fate decisions, very little is known about the topology, dynamics, or functional significance. Using human blood stem cell cultures as an experimental paradigm, we present a novel bioinformatic approach to integrate genome-scale molecular profiles (transcriptome and secretome) and publicly available databases to reconstruct soluble factor-mediated inter-cellular signalling networks regulating blood stem cell fate decisions. Umbilical cord blood UCB cells were lineage depleted via immuno-magnetic column and placed in serum free media, supplemented with SCF, FL, and TPO. At culture day 4, cells were either subject to Lin- cell re-selection + media exchange (S/E) or not (NS/NE) and cultured for an additional 4 days. At day-8, S/E cell sample was subject to another S/E procedure and cultured for an additional 4 days. At day-0, 4, 8, and 12, cells were separated into Lin- and Lin+ for independent profiling using Affymetrix HU133 v2.0 arrays.