scholarly journals THE SEPARATION OF DIFFERENT CELL CLASSES FROM LYMPHOID ORGANS

1969 ◽  
Vol 42 (3) ◽  
pp. 783-793 ◽  
Author(s):  
Ken Shortman ◽  
Kathrin Seligman
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Buoyant Density ◽  
Direct Consequence ◽  
Erythroid Cell ◽  
Mouse Bone Marrow ◽  
Erythroid Cells ◽  
Neutral Ph ◽  
Density Peaks ◽  
Density Shift

1. Mammalian erythrocytes swell as the pH of the isotonic suspending medium is lowered, as a direct consequence of the specialized permeability properties of the erythrocyte membrane. Lymphocytes and granulocytes from a variety of sources did not exhibit this property. 2. The behaviour of mouse bone marrow erythroid cells at various stages of differentiation was studied by using a change in buoyant density with pH as an index of swelling. The ability to swell with a pH drop was acquired while the cell was still nucleated. All non-nucleated cells showed swelling. Most small erythroblasts shared this property, whereas most large erythroblasts did not. 3. The density shift with pH was used to provide a purification scheme specific for erythroid cells. The bone marrow cells were first centrifuged to equilibrium in an isotonic albumin density gradient at neutral pH. Regions of the gradient containing the erythroid cells were collected, and the cells were recovered and redistributed in an albumin gradient at acid pH. The erythroid cells showed a specific density shift which removed them from contaminants. Preparations containing 90–97% erythroblasts were obtained by this technique. 4. Differentiation within the erythroid series was accompanied by a general increase in cell buoyant density at neutral pH. This density increase may have been a discontinuous process, since erythroid cells appeared to form a number of density peaks. 5. The pH shift technique, in association with established density distribution and sedimentation velocity procedures, provides a range of cell separation techniques for biological or biochemical studies of erythroid cell differentiation in the complex cell mixtures in bone marrow or spleen.

scholarly journals Cell interactions influencing murine marrow megakaryocytes: nature of the potentiator cell in bone marrow

Blood ◽  
1981 ◽  
Vol 57 (1) ◽  
pp. 157-163 ◽  
Author(s):  
N Williams ◽  
H Jackson ◽  
P Ralph ◽  
I Nakoinz
Keyword(s):  
Bone Marrow ◽  
Conditioned Medium ◽  
Colony Formation ◽  
Bone Marrow Cells ◽  
Buoyant Density ◽  
Colony Growth ◽  
Mouse Bone Marrow ◽  
Bone Marrow Cultures ◽  
Cell Conditioned Medium ◽  
Marrow Cells

Abstract Auxiliary bone marrow cells are required for optimal murine megakaryocyte colony formation in addition to progenitor cells and a colony stimulating activity (CSA) present in WEHI-3 cell conditioned medium. These auxiliary cells are adherent, with a sedimentation rate of 5.8 mm hr-1 and buoyant density of 1.065–1.078 gcm-3. The activity from bone marrow cells is loss at irradiation doses above 900 rad. Bone marrow cells with these characteristics, and supernatants from lung, bone shafts, and peritoneal exudate cells were all active in enhancing megakaryocyte colony incidences in mouse bone marrow cultures above those stimulated by an obligatory activity in WEHI-3 cell conditioned medium. Certain macrophage cell lines (J774, P388D1) could elaborate the activity. This study confirms that a potentiation activity enhances CSA stimulation of megakaryocyte colony formation. The potentiator is elaborated by bone marrow cells in limiting amounts requiring either high cell concentrations or an exogenous source of the activity for optimal colony growth.

scholarly journals Cyclic AMP phosphodiesterase activity during differentiation of rabbit erythroid bone marrow cells

1981 ◽  
Vol 196 (3) ◽  
pp. 887-892 ◽  
Author(s):  
M S Setchenska ◽  
H R Arnstein ◽  
J G Vassileva-Popova
Keyword(s):  
Bone Marrow ◽  
Cell Division ◽  
Cyclic Amp ◽  
Bone Marrow Cells ◽  
Velocity Sedimentation ◽  
Erythroid Cell ◽  
Erythroid Cells ◽  
Phosphodiesterase Activity ◽  
Physiological Control ◽  
Cyclic Amp Phosphodiesterase

Changes in the activity of cyclic AMP phosphodiesterase during differentiation of rabbit bone marrow erythroid cells were investigated. The cells were separated by velocity sedimentation at unit gravity into six fractions corresponding to different stages of development: proerythroblasts, basophilic cells, polychromatic cells, early orthochromatic and late orthochromatic cells and reticulocytes. Cyclic AMP phosphodiesterase was found to be very active in the most immature cells, the proerythroblasts, which also have the highest content of cyclic AMP. After differentiation into basophilic erythroblasts, a 4-fold decrease in cyclic AMP phosphodiesterase activity was observed. In these cells the amount of cyclic AMP was about 80% lower than that in proerythroblasts. In polychromatic cells a further drop in phosphodiesterase activity occurred. After the final cell division the enzyme activity was very low and the levels of cyclic AMP in the early and late orthochromatic cells remained constant. Kinetic studies demonstrated a heterogeneity of erythroid cell cyclic AMP phosphodiesterase: high affinity, low-Km (5.5 } 10(-6) M) and low affinity, high-Km (0.1 } 10(-3) M) enzymes were found. The phosphodiesterase activity was dependent on the presence of Mg2+ and was activated by Ca2+ at low Mg2+ concentrations (1 mM). The changes in cyclic AMP phosphodiesterase activity during differentiation and maturation of erythroid cells suggest the possible importance of this enzyme in the physiological control of cyclic AMP concentrations in developing erythroblasts. The loss of cyclic AMP phosphodiesterase activity after cessation of cell division supports the concept of the significance of the final cell division in erythroblast differentiation.

scholarly journals Cell interactions influencing murine marrow megakaryocytes: nature of the potentiator cell in bone marrow

Blood ◽  
1981 ◽  
Vol 57 (1) ◽  
pp. 157-163
Author(s):  
N Williams ◽  
H Jackson ◽  
P Ralph ◽  
I Nakoinz
Keyword(s):  
Bone Marrow ◽  
Conditioned Medium ◽  
Colony Formation ◽  
Bone Marrow Cells ◽  
Buoyant Density ◽  
Colony Growth ◽  
Mouse Bone Marrow ◽  
Bone Marrow Cultures ◽  
Cell Conditioned Medium ◽  
Marrow Cells

Auxiliary bone marrow cells are required for optimal murine megakaryocyte colony formation in addition to progenitor cells and a colony stimulating activity (CSA) present in WEHI-3 cell conditioned medium. These auxiliary cells are adherent, with a sedimentation rate of 5.8 mm hr-1 and buoyant density of 1.065–1.078 gcm-3. The activity from bone marrow cells is loss at irradiation doses above 900 rad. Bone marrow cells with these characteristics, and supernatants from lung, bone shafts, and peritoneal exudate cells were all active in enhancing megakaryocyte colony incidences in mouse bone marrow cultures above those stimulated by an obligatory activity in WEHI-3 cell conditioned medium. Certain macrophage cell lines (J774, P388D1) could elaborate the activity. This study confirms that a potentiation activity enhances CSA stimulation of megakaryocyte colony formation. The potentiator is elaborated by bone marrow cells in limiting amounts requiring either high cell concentrations or an exogenous source of the activity for optimal colony growth.

Binding of Transferrin and Uptake of Iron by Rat Erythroid Cells in Vitro

1977 ◽  
Vol 52 (1) ◽  
pp. 87-96 ◽  
Author(s):  
N. J. Verhoef ◽  
P. J. Noordeloos
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cell ◽  
Binding Sites ◽  
Bone Marrow Cells ◽  
Specific Binding ◽  
Marrow Cell ◽  
Erythroid Cell ◽  
Erythroid Cells ◽  
Low Concentrations ◽  
Rat Bone

1. The binding of transferrin and the uptake of iron by rat bone-marrow-cell suspensions was investigated by the use of transferrin doubly labelled with 125I and 59Fe. 2. The pattern of transferrin binding was found to depend on the transferrin concentration in the incubation medium. At relatively low concentrations, binding of transferrin at 0–4 °C was lower than the binding at 37°C. At higher concentrations no difference could be observed between binding at 0–4°C and at 37°C. This phenomenon was explained in terms of a rapid non-specific adsorption of transferrin at 0–4°C, which takes place especially at higher transferrin concentrations, and a specific binding of transferrin at 37°C observed presumably at low concentrations. 3. The maximum number of specific transferrin-binding sites was found to be approximately 190 000 sites per rat reticulocyte and 330 000 sites per nucleated rat bone-marrow cell. The latter number corresponds to 500 000–700 000 sites per nucleated erythroid cell. 4. It was concluded that maturation of the erythroid cell is accompanied with a progressive loss of transferrin binding sites on the cell membrane. 5. When bone-marrow cells obtained after incubation with doubly-labelled transferrin were lysed with distilled water or with the detergent Nonidet P-40, differences in the subcellular distribution of the radioactivities could be observed. 6. It was concluded that the membrane fraction contains appreciable amounts of 59Fe not bound to 125I-labelled transferrin, which indicates that dissociation of the iron—transferrin complex is one of the earlier steps in the mechanism of iron uptake by erythroid cells.

Jun N-terminal kinase promotes proliferation of immature erythroid cells and erythropoietin-dependent cell lines

Blood ◽  
2004 ◽  
Vol 104 (3) ◽  
pp. 696-703 ◽  
Author(s):  
Sarah M. Jacobs-Helber ◽  
Stephen T. Sawyer
Keyword(s):  
Cell Lines ◽  
Bone Marrow Cells ◽  
Dominant Negative ◽  
Mitogen Activated Protein Kinase ◽  
Erythroid Cell ◽  
Mouse Bone Marrow ◽  
Erythroid Cells ◽  
Erythroid Progenitors ◽  
Primary Mouse ◽  
Dependent Cell

AbstractErythropoietin (EPO) is the hormone necessary for development of erythrocytes from immature erythroid cells. EPO activates Jun N-terminal kinase (JNK), a member of the mitogen-activated protein kinase (MAPK) family in the EPO-dependent murine erythroid HCD57 cells. Therefore, we tested if JNK activity supported proliferation and/or survival of these cells. Treatment with the JNK inhibitor SP600125 inhibited JNK activity and EPO-dependent proliferation of HCD57 cells and the human EPO-dependent cell lines TF-1 and UT7-EPO. SP600125 also increased the fraction of cells in G2/M. Introduction of a dominant-negative form of JNK1 inhibited EPO-dependent proliferation in HCD57 cells but did not increase the fraction of cells in G2/M. Constitutive JNK activity was observed in primary murine erythroid progenitors. Treatment of primary mouse bone marrow cells with the SP600125 inhibitor reduced the number of erythroid burst-forming units (BFU-e's) but not the more differentiated erythroid colony-forming units (CFU-e's), and SP600125 protected the BFU-e's from apoptosis induced by cytosine arabinoside, demonstrating that the SP600125 inhibited proliferation of the BFU-e's. Therefore, JNK activity appears to be an important regulator of proliferation in immature, primary erythroid cells and 3 erythroid cell lines but may not be required for the survival or proliferation of CFU-e's or proerythroblasts.

The structure of pre-mRNA and mRNA from erythroid cells

1978 ◽  
Vol 36 (2) ◽  
pp. 234-235
Author(s):  
A.-M. Ladhoff ◽  
B.J. Thiele ◽  
Ch. Coutelle ◽  
S. Rosenthal
Keyword(s):  
Bone Marrow ◽  
Structural Transformation ◽  
Electron Micrographs ◽  
Ammonium Acetate ◽  
Bone Marrow Cells ◽  
Erythroid Cells ◽  
Globin Mrna ◽  
Ordered Structures ◽  
Rabbit Bone ◽  
Rabbit Reticulocytes

The suggested precursor-product relationship between the nuclear pre-mRNA and the cytoplasmic mRNA has created increased interest also in the structure of these RNA species. Previously we have been published electron micrographs of individual pre-mRNA molecules from erythroid cells. An intersting observation was the appearance of a contour, probably corresponding to higher ordered structures, on one end of 10 % of the pre-mRNA molecules from erythroid rabbit bone marrow cells (Fig. 1A). A virtual similar contour was observed in molecules of 9S globin mRNA from rabbit reticulocytes (Fig. 1B). A structural transformation in a linear contour occurs if the RNA is heated for 10 min to 90°C in the presence of 80 % formamide. This structural transformation is reversible when the denatured RNA is precipitated and redissolved in 0.2 M ammonium acetate.

scholarly journals Luteolin transforms the polarity of bone marrow-derived macrophages to regulate the cytokine storm

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Shuxia Wang ◽  
Shuhang Xu ◽  
Jing Zhou ◽  
Li Zhang ◽  
Xiaodong Mao ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Inflammatory Responses ◽  
Bone Marrow Cells ◽  
Membrane Surface ◽  
Inflammatory Factors ◽  
Mouse Bone Marrow ◽  
Macrophage Polarisation ◽  
Ifn Γ ◽  
Anti Inflammatory ◽  
M1 Type

Abstract Background Macrophages are indispensable regulators of inflammatory responses. Macrophage polarisation and their secreted inflammatory factors have an association with the outcome of inflammation. Luteolin, a flavonoid abundant in plants, has anti-inflammatory activity, but whether luteolin can manipulate M1/M2 polarisation of bone marrow-derived macrophages (BMDMs) to suppress inflammation is still unclear. This study aimed to observe the effects of luteolin on the polarity of BMDMs derived from C57BL/6 mice and the expression of inflammatory factors, to explore the mechanism by which luteolin regulates the BMDM polarity. Methods M1-polarised BMDMs were induced by lipopolysaccharide (LPS) + interferon (IFN)-γ and M2-polarisation were stimulated with interleukin (IL)-4. BMDM morphology and phagocytosis were observed by laser confocal microscopy; levels of BMDM differentiation and cluster of differentiation (CD)11c or CD206 on the membrane surface were assessed by flow cytometry (FCM); mRNA and protein levels of M1/M2-type inflammatory factors were performed by qPCR and ELISA, respectively; and the expression of p-STAT1 and p-STAT6 protein pathways was detected by Western-blotting. Results The isolated mouse bone marrow cells were successfully differentiated into BMDMs, LPS + IFN-γ induced BMDM M1-phenotype polarisation, and IL-4 induced M2-phenotype polarisation. After M1-polarised BMDMs were treated with luteolin, the phagocytosis of M1-polarized BMDMs was reduced, and the M1-type pro-inflammatory factors including IL-6, tumour necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), and CD86 were downregulated while the M2-type anti-inflammatory factors including IL-10, IL-13, found in inflammatory zone (FIZZ)1, Arginase (Arg)1 and CD206 were upregulated. Additionally, the expression of M1-type surface marker CD11c decreased. Nevertheless, the M2-type marker CD206 increased; and the levels of inflammatory signalling proteins phosphorylated signal transducer and activator of transcription (p-STAT)1 and p-STAT6 were attenuated and enhanced, respectively. Conclusions Our study suggests that luteolin may transform BMDM polarity through p-STAT1/6 to regulate the expression of inflammatory mediators, thereby inhibiting inflammation. Naturally occurring luteolin holds promise as an anti-inflammatory and immunomodulatory agent.

scholarly journals Brahmarasayana protects against Ethyl methanesulfonate or Methyl methanesulfonate induced chromosomal aberrations in mouse bone marrow cells

2012 ◽  
Vol 12 (1) ◽  
Author(s):  
Kanive Parashiva Guruprasad ◽  
Advait Subramanian ◽  
Vikram Jeet Singh ◽  
Raghavendra Sudheer Kumar Sharma ◽  
Puthiya Mundyat Gopinath ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chromosomal Aberrations ◽  
Bone Marrow Cells ◽  
Ethyl Methanesulfonate ◽  
Methyl Methanesulfonate ◽  
Mouse Bone Marrow ◽  
Mouse Bone Marrow Cells ◽  
Marrow Cells

scholarly journals Hepatocyte-like cells from directed differentiation of mouse bone marrow cells in vitro1

2005 ◽  
Vol 26 (4) ◽  
pp. 469-476 ◽  
Author(s):  
Xiao-lei SHI ◽  
Yu-dong QIU ◽  
Qiang LI ◽  
Ting XIE ◽  
Zhang-hua ZHU ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Mouse Bone Marrow ◽  
Directed Differentiation ◽  
Mouse Bone Marrow Cells ◽  
Marrow Cells

Induction of micronuclei in mouse bone marrow cells: An evaluation of nucleoside analogues used in the treatment of AIDS

1991 ◽  
Vol 18 (3) ◽  
pp. 168-183 ◽  
Author(s):  
Marcia D. Phillips ◽  
Bruce Nascimbeni ◽  
Raymond R. Tice ◽  
Michael D. Shelby ◽  
A. A. Van Zeeland
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Nucleoside Analogues ◽  
Mouse Bone Marrow ◽  
Mouse Bone Marrow Cells ◽  
Marrow Cells
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