Characterization of human bone marrow fibroblast colony-forming cells (CFU-F) and their progeny.

The bone marrow stromal cell population is comprised of fibroblasts, endothellal cells, fat cells and “reticular cells.” The characteristics and the regulatory role in hematopolesis of each of these cell types are unknown. A liquid culture system has been used to clone and to characterize human bone marrow fibroblast colony-forming cells (CFU-F). The linear relationship between the number of cells plated and the number of colonies formed suggests that fibroblastcolonies originate in a single cell. Bone marrow CFU-F were adherent and nonphagocytlc. The majority (90% ± 2%) were less dense than 1.070 g/cu cm. Velocity sedimentation separation demonstrated a heterogeneous CFU-F sedimentation rate, with a modal sedimentation of 4.95 ± 0.15 mm/hr. Analysisof CFU-F proliferative status by the thymidine suicide technique indicated that this cell was noncycling in individuals with undisturbed bone marrow function. Some of the more distinctive products of fibroblasts, other stromal cells, and hematopoietic colony-forming cells were used as positive and negative markers for CFU-F and the cells derived from them In vitro. Complement-mediated cytotoxicity using anti-la and anti-factor-VIll antigen antisera did not inhibit fibroblast colony formation. In contrast, a striking reduction of granulocyte-macrophage colony formation (CFU-c) was seen when bone marrow cells were treated with anti-la antiserum. Immunofluorescence staining was used to characterize the cells derived from CFU-F in vitro. No staining was observed after incubation of subconfluent cultures with anti-la and anti-factor-VIll antigen antisera. A positIve lmmunofluorescent staining was obtained when isolated antibodies against three of the main proteins of bone marrow matrix: type I collagen, type Ill collagen, and fibronectin were used. Ultrastructural analysis showed that CFU-F progeny, in contrast to endothellal cells, did not contain Welbel-Palade bodies. These data support the conclusion that the colonies described in this study are of fibroblastic nature.

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