Multiparameter Analysis of Human Bone Marrow Stromal Cells Identifies Distinct Immunomodulatory and Differentiation-Competent Subtypes

• Published in: Stem cell reports 2015-06, Vol.4 (6), p.1004-1015
• Main Authors: James, Sally, Fox, James, Afsari, Farinaz, Lee, Jennifer, Clough, Sally, Knight, Charlotte, Ashmore, James, Ashton, Peter, Preham, Olivier, Hoogduijn, Martin, Ponzoni, Raquel De Almeida Rocha, Hancock, Y., Coles, Mark, Genever, Paul
• Format: Article
• Language: English
• Subjects: Antigens, CD - metabolism; Bone Marrow Cells - cytology; Cell Differentiation; Cell Lineage; Cell Tracking; Cells, Cultured; Chemokine CXCL12 - metabolism; Cluster Analysis; GPI-Linked Proteins - metabolism; Humans; Interleukin-7 - metabolism; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - metabolism; Phenotype; Principal Component Analysis; Spectrum Analysis, Raman; Telomerase - genetics; Telomerase - metabolism; Transcriptome
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2015.05.005

Bone marrow stromal cells (BMSCs, also called bone-marrow-derived mesenchymal stromal cells) provide hematopoietic support and immunoregulation and contain a stem cell fraction capable of skeletogenic differentiation. We used immortalized human BMSC clonal lines for multi-level analysis of functional markers for BMSC subsets. All clones expressed typical BMSC cell-surface antigens; however, clones with trilineage differentiation capacity exhibited enhanced vascular interaction gene sets, whereas non-differentiating clones were uniquely CD317 positive with significantly enriched immunomodulatory transcriptional networks and high IL-7 production. IL-7 lineage tracing and CD317 immunolocalization confirmed the existence of a rare non-differentiating BMSC subtype, distinct from Cxcl12-DsRed+ perivascular stromal cells in vivo. Colony-forming CD317+ IL-7hi cells, identified at ∼1%–3% frequency in heterogeneous human BMSC fractions, were found to have the same biomolecular profile as non-differentiating BMSC clones using Raman spectroscopy. Distinct functional identities can be assigned to BMSC subpopulations, which are likely to have specific roles in immune control, lymphopoiesis, and bone homeostasis. •Immortalized BMSC clones were generated for in-depth functional and biophysical analysis•Distinct differentiation-competent and incompetent BMSC subsets are defined•Trilineage-competent BMSC clones exhibit enhanced vascular interaction gene sets•A non-differentiating, “immune-primed,” CD317+/IL-7hi BMSC subset was identified in vivo Genever and colleagues used immortalized human bone marrow stromal cell (BMSC) clones for in-depth analysis of subtype variation. They identified a rare, colony-forming BMSC subtype that lacked typical trilineage differentiation capacity, which under basal conditions displayed a marked immunomodulatory gene set, with high IL-7 and CD317 expression in vitro and in vivo.