Hematopoietic microenvironment. Origin, lineage, and transplantability of the stromal cells in long-term bone marrow cultures from chimeric mice.

Abstract

Studies of bone marrow transplant patients have suggested that the stromal cells of the in vitro hematopoietic microenvironment are transplantable into conditioned recipients. Moreover, in patients with myeloproliferative disorders, all of the stromal cells, which include presumptive endothelial cells, appear to be derived from hematopoietic precursors. To confirm these findings, we have constructed two chimeric mouse models: (a) traditional radiation chimeras, and (b) fetal chimeras, produced by placental injection of bone marrow into genetically anemic WX/WV fetuses, a technique that essentially precludes engraftment of nonhematopoietic cells. Using two-color indirect immunofluorescence, the stromal cells in long-term bone marrow culture derived from these chimeras were analyzed for donor or host origin by strain-specific H-2 antigens, and for cell lineage by a variety of other specific markers. 75-95% of the stromal cells were shown to be hematopoietic cells of the monocyte-macrophage lineage, based upon donor origin, phagocytosis, and expression of specific hematopoietic surface antigens. The remaining 5-25% of the stromal cells were exclusively host in origin. Apart from occasional fat cells, these cells uniformly expressed collagen type IV, laminin, and a surface antigen associated with endothelial cells. Since these endothelial-like cells are not transplantable into radiation or fetal chimeras, they are not derived from hematopoietic stem cells. The contrast between our findings and human studies suggests either unexpected species differences in the origin of stromal lineages or limitations in the previous methodology used to detect nonhematopoietic stromal cells.

Origin of marrow stromal cells and haemopoietic chimaerism following bone marrow transplantation determined by in situ hybridisation

“…It is generally believed that these two systems are distinct and do not arise from a common pluripotent stem cell (Owen, 1985). Evidence for this has come largely from analysis of adherent 'stromal' cells in long-term cultures of bone marrow derived from allogeneic marrow transplant recipients in both animals (Friedenstein et al, 1978;Bentley et al, 1982;Chertkov et al, 1985;Perkins & Fleischman, 1988) and humans (Golde et al, 1980;Laver et al, 1987;Simmons et al, 1987). However, similar studies in both humans (Keating et al, 1982;Piersma et al, 1983) and mice (Marshall et al, 1984) (Fialkow et al, 1980;Fialkow, 1983;Singer et al, 1984).…”

“…One of the major limiting factors with these in vitro models is the uncertainty about the nature and origin of adherent stromal cells which form in long-term marrow cultures (Dexter, 1982;Tavassoli & Friedenstein, 1983 (Perkins & Fleischman, 1988 (Greenberger, 1986).…”

“…Support for this concept has come from both cytogenetic and enzyme marker studies of adherent stromal cells in long-term marrow cultures derived from allogeneic transplant recipients (Keating et al, 1982;Piersma et al, 1983;Marshall et al, 1984). However, results of similar in vitro experiments in both animals (Friedenstein et al, 1978;Bentley et al, 1982;Chertkov et al, 1985;Perkins & Fleischman, 1988) and humans (Golde et al, 1980;Laver et al, 1987;Simmons et al, 1987) have shown a host origin for stromal cells, suggesting that a common precursor cell does not exist for both systems.…”

“…It is not possible to characterise fully and identify the cells which form the adherent cell population in these cultures, and it has been shown that they do contain cells of haemopoietic origin (Tavassoli & Friedenstein, 1983;Perkins & Fleischman, 1988). In this study, we have addressed this problem by examining intact bone marrow trephines without cell dissociation or culture.…”

Protection of committed murine haemopoietic progenitors against BCNU toxicity does not predict protection of primitive, multipotent spleen colony-forming cells – implications for chemoprotective gene therapy

“…However, the cumulative data published thus far suggest that the endothelial and fibroblast populations are likely to be of host origin. [69][70][71] Moreover, in the mice used in these experiments, reconstitution of the bone marrow was achieved using the non-adherent fraction of cells collected following co-cultivation with retroviral producers. It would seem unlikely, therefore, that many (adherent) fibroblast or endothelial cells were transferred and that this portion of the microenvironment would be host derived.…”