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Abstract
Spleen stromal lines which support in vitro hematopoiesis are investigated for their lineage origin and hematopoietic support function in vivo. Marker expression and gene profiling identify a lineage relationship with mesenchymal stem cells and perivascular reticular cells described recently in bone marrow. Stromal lines commonly express Cxcl12, Pdgfra and Pdgfr typical of bone marrow derived perivascular reticular cells but reflect a unique cell type in terms of other gene and marker expression. Their classification as osteoprogenitors is confirmed through ability to undergo osteogenic, but not adipogenic or chondrogenic differentiation. Some stromal lines were shown to form ectopic niches for HSCs following engraftment under the kidney capsule of NOD/SCID mice. The presence of myeloid cells and a higher representation of a specific dendritic-like cell type over other myeloid cells within grafts was consistent with previous in vitro evidence of hematopoietic support capacity. These studies reinforce the role of perivascular/perisinusoidal reticular cells in hematopoiesis and implicate such cells as niches for hematopoiesis in spleen.
Highlights
Data Availability Statement: All gene profiling data are available from the ArrayExpress database
Splenic stroma which support in vitro hematopoiesis reflect perivascular reticular cells. In line with their splenic origin, stromal lines STX3, 5G3, 3B5, 7G10 and 2A8 support myelopoiesis for an extended period when overlaid with splenocytes depleted of lymphoid and erythroid cells
Splenic stromal cell lines described here reflect mesenchymal perivascular and perisinusoidal reticular cells previously characterized as a heterogeneous population in bone marrow expressing PDGFRA/B, SCF and CXCL12 [71]
Summary
Data Availability Statement: All gene profiling data are available from the ArrayExpress database (accession number E-MTAB-8345). While evidence for HSC niches in spleen is increasing, very little is known about the stromal cells which support hematopoiesis and the hematopoietic cells which are produced, and whether these represent the full spectrum of blood cells, or restricted hematopoietic cell development Studies from this lab have identified a role for spleen in the production of a distinct dendritic-like cell type ‘L-DC’ which has been detected in both human and murine spleen [9,10,11]. Splenic stromal lines overlaid with hematopoietic progenitors, support long-term in vitro cultures with production of distinct myeloid cells [13]. A number of splenic stromal lines were isolated from long-term spleen cultures which were subsequently shown to support hematopoiesis in overlaid bone marrow progenitors [18, 19]. Stromal lines are for their ability to form an ectopic niche in vivo as a more physiological test of hematopoietic support capacity
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