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  • 1
    Electronic Resource
    Electronic Resource
    Production of colony stimulating factor in long-term bone marrow cultures (1983)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 114 (1983), S. 88-92 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Previous studies have shown no detectable colony-stimulating factor (CSF) in media harvested from long-term bone marrow cultures. In the present experiments supernatants from long-term cultures established in three laboratories were assayed for CSF by colony assay and by radioimmunoassay (RIA). Most samples were devoid of biologic activity but all contained CSF as judged by RIA. Biologic activity was found in the majority of samples after diafiltration to remove low molecular weight inhibitors or 5-fold concentration by ultrafiltration. Samples that remained inactive in the colony assay were subjected to gel filtration on Sephadex G-150 to remove potential high molecular weight inhibitors. Biologic activity remained lower than that by RIA in two of three samples tested. Thus, most long-term cultures appear to contain biologically active CSF but this activity is masked by various types of inhibitors. In addition some media appear to contain material that is only detected by RIA.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041140115
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  • 2
    Electronic Resource
    Electronic Resource
    Biochemical and functional characterization of proteoglycans produced by SI/SId murine bone marrow stromal cell lines (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 145 (1990), S. 53-59 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The Steel anemia of mice results from an inherited defect in the hematopoietic microenvironment. Proteoglycans synthesized by bone marrow stromal cells are an important functional component of the hematopoietic microenvironment in normal animals. It is thus possible that Steel anemia results from a molecular abnormality involving bone marrow stromal proteoglycans. To investigate this possibility, we studied proteoglycan synthesis in three stromal cell lines from Steel anemic (SI/SId) animals and two control stromal cell lines, one (+/+2.4) from a non-anemic littermate, and one (GBI/6) from a normal mouse. Proteoglycans were precursor labelled with 35S sulfate and separated by ion exchange HPLC, CsCI density gradient centrifugation, and molecular sieve HPLC. Glycosaminoglycan (GAG) moieties were characterized by molecular sieve HPLC and enzyme sensitivity. There were no consistent differences in total proteoglycan synthesis, proteoglycan heterogeneity, GAG hydrodynamic size, or enzyme sensitivity among the cell lines studied. Growth factor binding to stromal extracellular matrix (ECM) was studied by co-culture of an IL-3-dependent cell line (FDC-P1) with cell-free ECM preparations from an SI/SId and a control (GBI/6) stromal cell line, with and without pre-incubation with IL-3. Cell-free ECM preparations from SI/SId and control cell lines supported FDC-P1 growth to an approximately equal extent after pre-incubation with II-3. FDC-P1 growth support by ECM preparations from both cell lines was also observed without IL-3 pre-incubation, although to a lesser extent, suggesting ECM binding of endogenous growth factors synthesized by the stromal cells.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041450109
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  • 3
    Electronic Resource
    Electronic Resource
    Osteocalcin promotes differentiation of osteoclast progenitors from murine long-term bone marrow cultures (1994)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 55 (1994), S. 190-199 
    Details
    ISSN: 0730-2312
    Keywords: osteoclast ; osteocalcin ; bone marrow ; differentiation ; resorption ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Murine long-term bone marrow cultures (LTBMCs) were used to generate hematopoietic cells free from marrow stromal cells. These progenitor cells were treated with GM-CSF (5 U/ml) with or without rat bone osteocalcin or rat serum albumin in either α-MEM with 2% heat-inactivated horse serum alone (α) or supplemented with 10% L-cell-conditioned medium (as a source of M-CSF) (L10). Few substrate-attached cells survived in basal α medium, but when treated with L10 medium or GM-CSF, they survived and proliferated. Osteocalcin did not significantly affect survival or proliferation. Subcultures of cells treated with GM-CSF had large numbers of multinucleated cells, more than half of which were tartrate-resistant acid phosphatase-positive (TRAP). Osteocalcin further promoted the development of TRAP-positive multinucleated cells; a dose of 0.7 μg/ml osteocalcin promoted osteoclastic differentiation by 60%. Using a novel microphotometric assay, we detected significantly more tartrate-resistant acid phosphatase activity in the osteocalcin plus GM-CSF group (75.6 ± 14.2) than in GM-CSF alone (53.3 ± 7.3). In the absence of M-CSF, GM-CSF stimulated tartrate-resistant acid phosphatase activity, but osteocalcin did not have an additional effect. These studies indicate that osteocalcin promotes osteoclastic differentiation of a stromal-free subpopulation of hematopoietic progenitors in the presence of GM-CSF and L-cell-conditioned medium. These results are consistent with the hypothesis that this bone-matrix constituent plays a role in bone resorption. © 1994 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240550206
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  • 4
    Electronic Resource
    Electronic Resource
    Self-renewal of factor-dependent hemopoietic progenitor cell-lines derived from long-term bone marrow cultures demonstrates significant mouse strain genotypic variation (1980)
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 13 (1980), S. 501-511 
    Details
    ISSN: 0091-7419
    Keywords: bone marrow cultures ; hemopoiesis in vitro ; mouse genotype ; factor-dependent cell lines ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Long-term bone marrow cultures established from C57Ks/J mice have been shown to spontaneously release endogenous ecotropic RNA type-C virus (retrovirus). C57Ks/J marrow cultures produced granulocyte-macrophage progenitor cells (GM-CFUc) and immature and mature granulocytes for over 45 weeks. In contrast, NIH Swiss mouse marrow cultures failed to release detectable ecotropic virus and generated GM-CFUc and granulocytes for 25-35 weeks and established WEHI-3 conditioned medium (CM) dependent cell lines in vitro and did not establish permanent cell lines. To determine whether viral and/or cellular genes regulated the longevity of C57Ks/J marrow cultures, groups of cultures were established from the marrow of (NIH-Swiss × C57Ks/J) F1 hybrid, F2 hybrid, and (NIH Swiss × C57Ks/J) X NIH Swiss backcross generations. Release of endogenous ecotropic virus was measured weekly in each culture as was the duration of production of immature granulocytic cells and GM-CFUc over a 58-week period. The results demonstrated a complex pattern of inheritance of longevity of long-term in vitro hemopoiesis. Increased longevity did not absolutely correlate with detectable replication of the C57Ks/J N-tropic virus.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jss.400130409
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