Thrombopoietin-Induced Stimulation of Megakaryocyte- Enriched Bone Marrow Cultures

1979 ◽  
Author(s):  
K. L. Kellar ◽  
B. L. Evatt ◽  
C. R. McGrath ◽  
R. B. Ramsey
Keyword(s):  
Bone Marrow ◽  
Dna Synthesis ◽  
Bone Marrow Cells ◽  
Rabbit Plasma ◽  
Bone Marrow Cultures ◽  
Plasma Fractions ◽  
Marrow Cultures ◽  
Stimulation Of

Liquid cultures of bone marrow cells enriched for megakaryocytes were assayed for incorporation of 3H-thymidine (3H-TdR) into acid-precipitable cell digests to determine the effect of thrombopoietin on DNA synthesis. As previously described, thrombopoietin was prepared by ammonium sulfate fractionation of pooled plasma obtained from thrombocytopenic rabbits. A control fraction was prepared from normal rabbit plasma. The thrombopoietic activity of these fractions was determined in vivo with normal rabbits as assay animals and the rate of incorporation of 75Se-selenomethionine into newly formed platelets as an index of thrombopoietic activity of the infused material. Guinea pig megakaryocytes were purified using bovine serum albumin gradients. Bone marrow cultures containing 1.5-3.0x104 cells and 31%-71% megakaryocytes were incubated 18 h in modified Dulbecco’s MEM containing 10% of the concentrated plasma fractions from either thrombocytopenic or normal rabbits. In other control cultures, 0.9% NaCl was substituted for the plasma fractions. 3H-TdR incorporation was measured after cells were incubated for 3 h with 1 μCi/ml. The protein fraction containing thrombopoietin-stimulating activity caused a 25%-31% increase in 3H-TdR incorporation over that in cultures which were incubated with the similar fraction from normal plasma and a 29% increase over the activity in control cultures to which 0.9% NaCl had been added. These data suggest that thrombopoietin stimulates DNA synthesis in megakaryocytes and that this tecnique may be useful in assaying thrombopoietin in vitro.

scholarly journals Cellular maturation in human preleukemia

Blood ◽  
1978 ◽  
Vol 52 (2) ◽  
pp. 355-361
Author(s):  
HP Koeffler ◽  
DW Golde
Keyword(s):  
Bone Marrow ◽  
Cell Differentiation ◽  
Liquid Culture ◽  
Cultured Cells ◽  
Bone Marrow Cells ◽  
Myelogenous Leukemia ◽  
Bone Marrow Cultures ◽  
Acute Myelogenous ◽  
Marrow Cultures

Bone marrow cells from three preleukemic patients with prominent marrow karyotypic abnormalities were studied in liquid culture to determine if the neoplastic clones were capable of maturation. Parallel cytogenetic and cytologic studies were performed in sequentially harvested bone marrow cultures. Maturation, albeit delayed, occurred in cultures from all three patients. By 14 days of culture in vitro, morphologic, cytochemical, and functional evidence of maturation was observed in about 70% of the cells. By day 21, 85% of the cells were mature by these criteria. All but 2 of 249 metaphases from the cultured cells contained the cytogenetic abnormality of the neoplastic clone. We conclude that some preleukemic cells identified by a chromosomal abnormality can mature in vitro. Preleukemia may be viewed as a syndrome of “early leukemia” in which the neoplastic clone is established and manifested functionally as ineffective hematopoiesis. Hematopoietic cell differentiation becomes progressively abnormal with termination in the nearly complete maturational block characteristic of acute myelogenous leukemia.

The detection of in vivo hematotoxicity of benzene by in vitro liquid bone marrow cultures

1981 ◽  
Vol 60 (2) ◽  
pp. 346-353 ◽  
Author(s):  
Kenichi Harigaya ◽  
Marilyn E. Miller ◽  
Eugene P. Cronkite ◽  
Robert T. Drew
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

STIMULATION OF BONE MARROW CELLS AND BONE FORMATION BY NACRE. IN VIVO AND IN VITRO STUDIES

Bioceramics ◽  
1999 ◽  
Author(s):  
M. Lamghari ◽  
S. Berland ◽  
A. Laurent ◽  
H. Huet ◽  
M.J. Almeida ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Formation ◽  
Bone Marrow Cells ◽  
In Vitro Studies ◽  
Stimulation Of ◽  
Marrow Cells

The anabolic effect of PGE2in rat bone marrow cultures is mediated via the EP4 receptor subtype

1999 ◽  
Vol 276 (2) ◽  
pp. E376-E383 ◽  
Author(s):  
M. Weinreb ◽  
A. Grosskopf ◽  
N. Shir
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Osteoblastic Differentiation ◽  
Nodule Formation ◽  
Prostaglandin E ◽  
Receptor Subtype ◽  
Ep4 Receptor ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

Prostaglandin E2(PGE2) is an anabolic agent in vivo that stimulates bone formation by recruiting osteoblasts from bone marrow precursors. To understand which of the known PGE2 receptors (EP1–4) is involved in this process, we tested the effect of PGE2 and various EP agonists and/or antagonists on osteoblastic differentiation in cultures of bone marrow cells by counting bone nodules and measuring alkaline phosphatase activity. PGE2increased both parameters, peaking at 100 nM, an effect that was mimicked by forskolin and was abolished by 2′,3′-dideoxyadenosine (an adenylate cyclase inhibitor) and was thus cAMP dependent, pointing to the involvement of EP2 or EP4. Consistently, 17-phenyl-ω-trinor PGE2(EP1 agonist) and sulprostone (EP3/EP1agonist) lacked any anabolic activity. Furthermore, butaprost (EP2 agonist) was inactive, 11-deoxy-PGE1(EP4/EP2agonist) was as effective as PGE2, and the PGE2 effect was abolished dose dependently by the selective EP4 antagonist AH-23848B, suggesting the involvement of EP4. We also found that PGE2 increased nodule formation and AP activity when added for the initial attachment period of 24 h only. Thus this study shows that PGE2 stimulates osteoblastic differentiation in bone marrow cultures, probably by activating the EP4 receptor, and that this effect may involve recruitment of noncommitted (nonadherent) osteogenic precursors, in agreement with its suggested mode of operation in vivo.

M-CSF: a novel plasmacytoid and conventional dendritic cell poietin

Blood ◽  
2008 ◽  
Vol 111 (1) ◽  
pp. 150-159 ◽  
Author(s):  
Ben Fancke ◽  
Mark Suter ◽  
Hubertus Hochrein ◽  
Meredith O'Keeffe
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
High Capacity ◽  
Interferon Α ◽  
Bone Marrow Cultures ◽  
A Cell ◽  
Macrophage Colony ◽  
Major Attention

The critical importance of plasmacytoid dendritic cells (pDCs) in viral infection, autoimmunity, and tolerance has focused major attention on these cells that are rare in blood and immune organs of humans and mice. The recent development of an Flt-3 ligand (FL) culture system of bone marrow cells has led to the simple generation of large numbers of pDCs that resemble their in vivo steady-state counterparts. The FL system has allowed unforeseen insight into the biology of pDCs, and it is assumed that FL is the crucial growth factor for these cells. Surprisingly we have found that a cell type with high capacity for interferon-α (IFN-α) production in response to CpG-containing oligonucleotides, a feature of pDCs, develop within macrophage–colony-stimulating factor (M-CSF)–generated bone marrow cultures. Analysis of this phenomenon revealed that M-CSF is able to drive pDCs as well as conventional DCs (cDCs) from BM precursor cells in vitro. Furthermore, application of M-CSF to mice was able to drive pDCs and cDCs development in vivo. It is noteworthy that using mice deficient in FL indicated that the M-CSF-driven generation of pDCs and cDCs in vitro and in vivo was independent of endogenous FL.

scholarly journals Multistep virus-induced leukemogenesis in vitro: description of a model specifying three steps within the myeloblastic malignant process.

1984 ◽  
Vol 4 (1) ◽  
pp. 216-220 ◽  
Author(s):  
J M Heard ◽  
S Fichelson ◽  
B Sola ◽  
M A Martial ◽  
B Varet ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Leukemia Virus ◽  
Latency Period ◽  
In Vitro Production ◽  
Long Latency ◽  
Permanent Cell ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

A helper-independent Friend leukemia virus was used to infect bone marrow cultures. This virus induces myeloblastic leukemia in mice after a long latency period. Infection of the bone marrow cultures resulted in the in vitro production of myeloblastic leukemogenesis after a long latency period. Three steps were observed in the evolution of the infected cultures, and permanent cell lines were derived at each step. This allowed us to individualize three successive events in the course of the myeloblastic transformation: (i) an abnormal responsiveness to the physiological hormone granulo-macrophagic colony-stimulating factor, (ii) the acquisition of growth autonomy, and (iii) the acquisition of in vivo tumorigenicity.

scholarly journals Cellular maturation in human preleukemia

Blood ◽  
1978 ◽  
Vol 52 (2) ◽  
pp. 355-361 ◽  
Author(s):  
HP Koeffler ◽  
DW Golde
Keyword(s):  
Bone Marrow ◽  
Cell Differentiation ◽  
Liquid Culture ◽  
Cultured Cells ◽  
Bone Marrow Cells ◽  
Myelogenous Leukemia ◽  
Bone Marrow Cultures ◽  
Acute Myelogenous ◽  
Marrow Cultures

Abstract Bone marrow cells from three preleukemic patients with prominent marrow karyotypic abnormalities were studied in liquid culture to determine if the neoplastic clones were capable of maturation. Parallel cytogenetic and cytologic studies were performed in sequentially harvested bone marrow cultures. Maturation, albeit delayed, occurred in cultures from all three patients. By 14 days of culture in vitro, morphologic, cytochemical, and functional evidence of maturation was observed in about 70% of the cells. By day 21, 85% of the cells were mature by these criteria. All but 2 of 249 metaphases from the cultured cells contained the cytogenetic abnormality of the neoplastic clone. We conclude that some preleukemic cells identified by a chromosomal abnormality can mature in vitro. Preleukemia may be viewed as a syndrome of “early leukemia” in which the neoplastic clone is established and manifested functionally as ineffective hematopoiesis. Hematopoietic cell differentiation becomes progressively abnormal with termination in the nearly complete maturational block characteristic of acute myelogenous leukemia.

Stimulation of bone marrow cells and bone formation by nacre: in vivo and in vitro studies

Bone ◽  
1999 ◽  
Vol 25 (2) ◽  
pp. 91S-94S ◽  
Author(s):  
M Lamghari ◽  
M.J Almeida ◽  
S Berland ◽  
H Huet ◽  
A Laurent ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Formation ◽  
Bone Marrow Cells ◽  
In Vitro Studies ◽  
Stimulation Of ◽  
Marrow Cells
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