Human bone marrow and umbilical cord-derived mesenchymal stromal cells activated with TNF-α, IL-1β or IFN-γ

We used microarrays to characterize the in vitro response of three populations each of neonatal and adult MSCs to three soluble inflammatory mediators separately, in xeno-free media. The magnitude of transcriptomic response to each priming condition was compared between MSC sources. Whether resting MSC source differences were attentuated or amplified following activation with cytokines was also explored. Cryopreserved MSCs were obtained from commercial sources: Wharton’s jelly (UC) MSCs from Tissue Regeneration Therapeutics, Inc., and BM MSCs from RoosterBio Inc.. Three populations of each MSC source (2 male and 1 female donors each) were expanded in xeno-free media, and at comparable population doubling levels, were seeded into 3 flasks each for priming. Media was replaced and purified recombinant human cytokines TNF-α, IL-1β or IFN-γ were added to each culture in protein-free media for 24 hours. Conditioned media was collected for quantification of secreted protein content, and RNA was extracted from the resting (unprimed) or primed cells and analyzed by Affymetrix Human Genome U133A 2.0 Array. Batches of experimental culture and GeneChip processing time frames were balanced to eliminate additional variability or bias in the data, and batch information is included.