GP130 and c-kit signalling, initiated by the sIL-6R/IL-6 complex, is insufficient to expand the primitive adult bone marrow CD34+CD38- pre-CFu cell

It has previously been shown that gp130 and c-kit signalling synergize for the ex vivo expansion of human cord blood (CB) CD34 + haematopoietic progenitor cells. We were interested in evaluating this synergy within an ontogenetically different haematopoietic tissue [i.e. adult bone marrow (BM)] and on a more primitive progenitor subset (i.e. CD34 + CD38 -cells), which are highly enriched for pre-colony forming unit (CFU) cells. These cells were plated out in a primary liquid culture supplemented with either interleukin (IL)-6+stem cell factor (SCF), IL-6+SCF+soluble IL-6 receptor (sIL-6R), IL-6+SCF+sIL-6R+IL-3+IL-1 or SCF+IL-3+IL-6+IL-1. Cell counting after liquid culture revealed an absolute expansion of 2.2-, 4.1-, 89.5- and 65.7-fold compared with initial cell input for the four-cytokine combinations, respectively. The secondary read-out assay revealed that this cell expansion in the liquid culture also resulted in CFU generation, with absolute cloning efficiencies of 0.002, 0.024, 12.13 and 7.73 (per cell initially present) for the respective cytokine combinations. These results indicate that gp130 and c-kit signalling alone (i.e. using IL-6+SCF+sIL-6R), in terms of both cell number and CFU generation, insufficiently stimulate primitive adult BM CD34+CD38- haematopoietic cells in order to reach a CFU generation comparable with that obtained after multifactor stimulation. Adding sIL-6R to the multifactor stimulation and compared with this multifactor stimulation, a 1.7-fold synergy in terms of cell expansion and a 3.0-fold synergy in terms of CFU generation are obtained. The sIL-6R/IL-6 complex thus has a narrower spectrum of action on primitive adult BM CD34+CD38 - cells than on CB CD34 + cells.