Role of cell contacts for differentiation of precursor cells of hematopoietic stroma in long-term bone marrow cultures

1982 ◽  
Vol 94 (2) ◽  
pp. 1128-1131
Author(s):  
O. A. Gurevich ◽  
N. I. Drize ◽  
I. L. Chertkov
Keyword(s):  
Bone Marrow ◽  
Precursor Cells ◽  
Cell Contacts ◽  
Bone Marrow Cultures ◽  
Hematopoietic Stroma ◽  
Marrow Cultures

Megakaryocytic maturation in murine long-term bone marrow culture: role of interleukin-6

Blood ◽  
1991 ◽  
Vol 78 (6) ◽  
pp. 1438-1447
Author(s):  
RL Mei ◽  
SA Burstein
Keyword(s):  
Bone Marrow ◽  
Interleukin 6 ◽  
Maturation In Vitro ◽  
Bone Marrow Cultures ◽  
Acetylcholinesterase Staining ◽  
Stromal Layer ◽  
Marrow Cultures

Megakaryocytic maturation was analyzed in long-term bone marrow cultures in the absence of added growth factors. Megakaryocytes could be observed for periods of up to 13 weeks in both the supernatant and stromal layer of these cultures. Using acetylcholinesterase staining for enumeration and sizing of megakaryocytes, and a novel rat antimurine platelet monoclonal antibody (MoAb) that detects only megakaryocytes in bone marrow, the number, volume, and ploidy of these cells were assessed microscopically and by flow cytometry. Correlation of these measurements with ambient interleukin-6 (IL-6) levels showed no relationship between IL-6 bioactivity and megakaryocyte number. Conversely, the relatively high IL-6 bioactivity present during the first 2 weeks of culture was correlated with increased megakaryocytic size and ploidy, while the relatively lower IL-6 bioactivity present after week 3 corresponded to decreased megakaryocytic size and ploidy. Addition of neutralizing anti-IL-6 MoAb decreased megakaryocytic size and ploidy at times when ambient IL-6 levels were relatively high, while the addition of exogenous IL-6 increased size and ploidy at times when endogenous IL-6 concentrations were low. The data show that long- term bone marrow cultures can be used as a means to evaluate megakaryocytic maturation in vitro, and suggest that, to some extent, IL-6 plays a role in the maturation process in this system.

scholarly journals Megakaryocytic maturation in murine long-term bone marrow culture: role of interleukin-6

Blood ◽  
1991 ◽  
Vol 78 (6) ◽  
pp. 1438-1447 ◽  
Author(s):  
RL Mei ◽  
SA Burstein
Keyword(s):  
Bone Marrow ◽  
Interleukin 6 ◽  
Maturation In Vitro ◽  
Bone Marrow Cultures ◽  
Acetylcholinesterase Staining ◽  
Stromal Layer ◽  
Marrow Cultures

Abstract Megakaryocytic maturation was analyzed in long-term bone marrow cultures in the absence of added growth factors. Megakaryocytes could be observed for periods of up to 13 weeks in both the supernatant and stromal layer of these cultures. Using acetylcholinesterase staining for enumeration and sizing of megakaryocytes, and a novel rat antimurine platelet monoclonal antibody (MoAb) that detects only megakaryocytes in bone marrow, the number, volume, and ploidy of these cells were assessed microscopically and by flow cytometry. Correlation of these measurements with ambient interleukin-6 (IL-6) levels showed no relationship between IL-6 bioactivity and megakaryocyte number. Conversely, the relatively high IL-6 bioactivity present during the first 2 weeks of culture was correlated with increased megakaryocytic size and ploidy, while the relatively lower IL-6 bioactivity present after week 3 corresponded to decreased megakaryocytic size and ploidy. Addition of neutralizing anti-IL-6 MoAb decreased megakaryocytic size and ploidy at times when ambient IL-6 levels were relatively high, while the addition of exogenous IL-6 increased size and ploidy at times when endogenous IL-6 concentrations were low. The data show that long- term bone marrow cultures can be used as a means to evaluate megakaryocytic maturation in vitro, and suggest that, to some extent, IL-6 plays a role in the maturation process in this system.

Formation of Multinucleated Cells that Respond to Osteotropic Hormones in Long Term Human Bone Marrow Cultures*

Endocrinology ◽  
1987 ◽  
Vol 120 (6) ◽  
pp. 2326-2333 ◽  
Author(s):  
B. R. MACDONALD ◽  
N. TAKAHASHI ◽  
L. M. MCMANUS ◽  
J. HOLAHAN, ◽  
G. R. MUNDY ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Multinucleated Cells ◽  
Bone Marrow Cultures ◽  
Human Bone Marrow Cultures ◽  
Marrow Cultures

scholarly journals Early pre-B-cell transformation induced by the v-fms oncogene in long-term mouse bone marrow cultures.

1989 ◽  
Vol 9 (9) ◽  
pp. 3973-3981 ◽  
Author(s):  
G V Borzillo ◽  
C J Sherr
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Chain Gene ◽  
Mouse Bone Marrow ◽  
Feeder Layers ◽  
Bone Marrow Cultures ◽  
B Lymphoid ◽  
Marrow Cultures

Murine long-term bone marrow cultures that support B-lymphoid-cell development were infected with a helper-free retrovirus containing the v-fms oncogene. Infection of B-lymphoid cultures resulted in the rapid clonal outgrowth of early pre-B cells, which grew to high cell densities on stromal cell feeder layers, expressed v-fms-coded glycoproteins, and underwent immunoglobulin heavy-chain gene rearrangements. Late-passage cultures gave rise to factor-independent variants that proliferated in the absence of feeder layers, developed resistance to hydrocortisone, and became tumorigenic in syngeneic mice. The v-fms oncogene therefore recapitulates known effects of the v-abl and bcr-abl oncogenes on B-lineage cells. The ability of v-fms to induce transformation of early pre-B cells in vitro underscores the capacity of oncogenic mutants of the colony-stimulating factor-1 receptor to function outside the mononuclear phagocyte lineage.

scholarly journals Proteoglycans in human long-term bone marrow cultures: biochemical and ultrastructural analyses

Blood ◽  
1986 ◽  
Vol 67 (5) ◽  
pp. 1333-1343 ◽  
Author(s):  
TN Wight ◽  
MG Kinsella ◽  
A Keating ◽  
JW Singer
Keyword(s):  
Bone Marrow ◽  
Extracellular Matrix ◽  
Cell Layer ◽  
Ruthenium Red ◽  
Adherent Cells ◽  
Sulfate Proteoglycan ◽  
Chondroitinase Abc ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

Proteoglycans within the extracellular matrix of human bone marrow have been implicated in the process of hematopoiesis, but little is known about the structure and composition of these macromolecules in this tissue. Hematopoietically active human long-term bone marrow cultures were incubated with medium containing 35S-sulfate and 3H-glucosamine as labeling precursors. Proteoglycans present in the medium and cell layer were extracted with 4 mol/L guanidine HCI and purified by diethylaminoethyl (DEAE)-Sephacel ion exchange and molecular sieve chromatography. Both culture compartments contain a large chondroitin sulfate proteoglycan (MI, CI) that eluted in the void volume of a Sepharose CL-4B column and contained glycosaminoglycan chains of molecular weight (mol wt) approximately 38,000. A second population of sulfate-labeled material was identified as a broad heterogenous peak (MII, CII) that was included on Sepharose CL-4B at Kav = 0.31. This material when chromatographed on Sepharose CL-6B could be further separated into a void peak (MIIa, CIIa) and an included peak eluting at Kav = 0.39 (MIIb, CIIb). The void peaks (MIIa, CIIa) were susceptible to chondroitinase ABC digestion (99%) but slightly less susceptible to chondroitinase AC digestion (90%). Papain digestion of these peaks revealed them to be proteoglycans with glycosaminoglycan chains of mol wt approximately 38,000. The included peaks on Sepharose CL-6B (MIIb, CIIb) from both medium and cell layer compartments resisted digestion with papain, indicating the presence of glycosaminoglycan chains of mol wt approximately 38,000 either free or attached to a small peptide. Although this material was susceptible to chondroitinase ABC (98%), it was considerably less susceptible to chondrotinase AC (approximately 60%), indicating that it contained dermatan sulfate. A small amount of heparan sulfate proteoglycan was also identified but constituted only approximately 10% of the total sulfated proteoglycan extracted from these cultures. Additionally, approximately 40% of the incorporated 3H- activity radioactivity was present as hyaluronic acid. Electron microscopy revealed a layer of adherent cells covered by a mat containing ruthenium red-positive granules that were connected by thin filaments. The extracellular matrix layer above the adherent cells contained a mixture of hematopoietic cells. Chondroitinase ABC treatment of the cultures completely removed the ruthenium red-positive granules overlying the cells and resulted in a loss of approximately 70% of the 35S-sulfate-labeled material from the cell layer.(ABSTRACT TRUNCATED AT 400 WORDS)

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