A novel hematopoietic multilineage clone, Myl-D-7, is stromal cell-dependent and supported by an alternative mechanism(s) independent of stem cell factor/c-kit interaction.
A strictly stroma-dependent hematopoietic clone, Myl-D-7, with lympho-myeloid potential has been isolated. A subset of cells expresses myeloid-macrophage (Mac-1 and Gr-1), erythroid (TER119), and lymphoid (Thy-1 and B220) lineage markers. Spontaneous differentiation to the myeloid-macrophage, erythroid, or lymphoid pathway can be seen by morphologic criteria, detection of beta major globin synthesis, or expression of the early lymphoid specific transcription factor, Ikaros. By sorting lineage marker (Mac-1, Gr-1, B220, and TER119)-negative (LIN-) cells, we showed that the LIN- population actively self-renews on top of MS-5 stromal cells, and differentiates to LIN+ cells. Removal of stroma induces apoptosis and none of the growth factors tested can prevent apoptosis. Granulocyte-macrophage colony-stimulating factor accelerates the differentiation towards the myeloid-macrophage lineage. Using this clone, we show that (1) contact with stroma induces expression of bcl-2, (2) stromal cells derived from SI/SI homozygous fetuses can support long-term growth, and (3) conditioned media of specific stromal cells contains an activity that supports proliferation and self-renewal of the clone. Myl-D-7 can thus be used as an indicator cell for unknown factors that may provide stromal cell support.
Itoh
K
K
Heinrich-Pette-Institute for Experimental Virology and Immunology, Hamburg University, Germany.
Friel
J
J
Kluge
N
N
Kina
T
T
Kondo-Takaori
A
A
Kawamata
S
S
Uchiyama
T
T
Ostertag
W
W
D000818:Animals; D017209:Apoptosis; D001685:Biological Factors / Q000502:physiology*; D015415:Biomarkers / Q000032:analysis; D001854:Bone Marrow Cells; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D019070:Cell Lineage; D002999:Clone Cells / Q000166:cytology; D018920:Coculture Techniques; D003114:Colony-Forming Units Assay; D003238:Connective Tissue / Q000502:physiology*; D003239:Connective Tissue Cells; D017077:Culture Media, Conditioned; D003673:Defective Viruses / Q000235:genetics; D005260:Female; D005622:Friend murine leukemia virus / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006378:Helper Viruses / Q000235:genetics; D016298:Hematopoietic Cell Growth Factors / Q000032:analysis / Q000235:genetics / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D051379:Mice; D008811:Mice, Inbred DBA; D008817:Mice, Mutant Strains; D016513:Mice, SCID; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2; D019009:Proto-Oncogene Proteins c-kit / Q000502:physiology; D017978:Receptors, Growth Factor / Q000032:analysis / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D009053:Sarcoma Viruses, Murine / Q000235:genetics; D013154:Spleen / Q000166:cytology; D013155:Spleen Focus-Forming Viruses / Q000235:genetics; D019089:Stem Cell Factor / Q000502:physiology
D001685:Biological Factors; D015415:Biomarkers; D017077:Culture Media, Conditioned; D016298:Hematopoietic Cell Growth Factors; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D017978:Receptors, Growth Factor; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D019009:Proto-Oncogene Proteins c-kit