Subpopulations of stromal cells from long-term human bone marrow cultures: ontogeny of progenitor cells and expression of growth hormone receptors.
Long-term culture of bone marrow derived stromal colony forming cells (S-CFC) in matrix and nutrient defined agar medium resulted in stromal cell colonies that pass sequentially through three distinct morphological stages: firstly, aggregated loose syncytium of round to avoid cells (stage I), a second developmental stage of large branching colonies in which the cells become enlarged, elongated with cytoplasmic projections forming a loosely anastomized network with adjacent cells (stage II), and finally cells become dissociated, loosing their long, thin cytoplasmic filaments and breaking their contacts with one another, but remain large and retain a bi-polar nature (stage III). Cells were also grown in liquid medium in a culture microenvironment closely resembling conditions of haemopoiesis in vitro. Using a panel of well defined monoclonal antibodies reactive against the rat, rabbit and human growth hormone receptors, this study found immunochemical evidence of the presence and localization of binding sites of growth hormone (GH) in the cell membrane and extra-nuclear Golgi area of long-term bone marrow derived human stromal cells in liquid and semi-solid nutrient agar mediums. GH-receptor immunoreactivity was present in small proliferating progenitor cells, myofibroblast-like cells, large reticular fibroblast cells, adipocytes and endothelial cells. Only MAb known to be reactive against human tissue resulted in strong immunoreactivity. The expression of GH-receptors not only on small proliferating, but also on the well differentiated cells, indicates a role for growth hormone on non-progenitor cells. GH-receptor immunoreactivity on differentiating and/or differentiated cells suggests that GH is also necessary for, or has a trophic function in differentiation. We propose that direct GH action is necessary not only for differentiation of progenitor cells as implied by the dual effector hypothesis, but also their subsequent clonal expansion, differentiation and maintenance.
Lincoln
D T
DT
Department of Medical Laboratory Technology, Faculty of Allied Health Sciences, Kuwait University, Kuwait.
Sinowatz
F
F
Gabius
S
S
Gabius
H J
HJ
Temmim
L
L
Baker
H
H
Mathew
T C
TC
Waters
M J
MJ
D000818:Animals; D000911:Antibodies, Monoclonal; D001853:Bone Marrow / Q000378:metabolism; D001854:Bone Marrow Cells; D002452:Cell Count; D002455:Cell Division; D002478:Cells, Cultured; D002648:Child; D006801:Humans; D007150:Immunohistochemistry; D011817:Rabbits; D051381:Rats; D011986:Receptors, Somatotropin / Q000096:biosynthesis*; D013234:Stem Cells / Q000378:metabolism / Q000648:ultrastructure*; D017154:Stromal Cells / Q000378:metabolism / Q000648:ultrastructure; D013997:Time Factors
D000911:Antibodies, Monoclonal; D011986:Receptors, Somatotropin