STUDIES ON THE CULTIVATION IN VITRO OF NORMAL AND PATHOLOGICAL GUINEA-PIG BONE MARROW

1946 ◽  
Vol 24 (2) ◽  
pp. 139-142
Author(s):  
L Brandon Fastier
Keyword(s):  
Bone Marrow ◽  
Guinea Pig ◽  
Cultivation In Vitro

scholarly journals Identity and Proliferation of Small Lymphocyte Precursors in Cultures of Lymphocyte-rich Fractions of Guinea Pig Bone Marrow

Blood ◽  
1971 ◽  
Vol 37 (1) ◽  
pp. 73-86 ◽  
Author(s):  
Y. YOSHIDA ◽  
D. G. OSMOND
Keyword(s):  
Bone Marrow ◽  
Guinea Pig ◽  
Lymphoid Cells ◽  
Short Term ◽  
Small Lymphocyte ◽  
Undifferentiated Cells ◽  
The Mean ◽  
High Concentrations ◽  
Transitional Cells

Abstract Radioautography with 3H-thymidine was used to examine the proliferative activity of bone marrow lymphoid cells and to identify the precursor cells of small lymphocytes in short-term cultures of lymphocyte-rich marrow fractions. High concentrations of small lymphocytes (nuclear diameters less than 8.0 µ in smears) together with large lymphoid ("transitional") cells (nuclear diameters greater than 8.0 µ) were separated from suspensions of guinea pig bone marrow by centrifugation in linear sucrose-serum density gradients. When such lymphocyte-rich marrow fractions were cultured in vitro the labeling and mitotic indices following either continuous or terminal exposure to 3H-thymidine indicated that the large lymphoid cells were confined mainly to the pre-DNA-synthetic (G1) and early DNA-synthetic (S) phases at first, but proceeded subsequently through S phase and mitosis. From these data tentative values were derived for the in vitro duration of G1 (12 hours) and S (13.7 hours). Further cultures were followed radioautographically after a 1-hour pulse of 3H-thymidine at 6-7 hours of culture. The absolute numbers of labeled large lymphoid cells declined during the subsequent 21 hours but, simultaneously, labeled small lymphocytes appeared and increased progressively in absolute numbers to 44.4 ± 8.1 per cent of the initial numbers of labeled large lymphoid cells. The mean grain count of labeled small lymphocytes was half that of the initially labeled large lymphoid cells. Very few labeled undifferentiated cells other than large lymphoid cells were observed. The results demonstrate that lymphocyte-rich marrow fractions are capable of sustaining the production of small lymphocytes in short-term cultures and that the immediate precursors of marrow small lymphocytes are contained within a population of large lymphoid cells.

scholarly journals Eotaxin Induces a Rapid Release of Eosinophils and Their Progenitors From the Bone Marrow

Blood ◽  
1998 ◽  
Vol 91 (7) ◽  
pp. 2240-2248 ◽  
Author(s):  
Roger T. Palframan ◽  
Paul D. Collins ◽  
Timothy J. Williams ◽  
Sara M. Rankin
Keyword(s):  
Bone Marrow ◽  
Guinea Pig ◽  
Allergic Inflammation ◽  
Interleukin 5 ◽  
In Situ Perfusion ◽  
Dose Dependent ◽  
Femoral Bone Marrow

The CC-chemokine eotaxin is a potent eosinophil chemoattractant that stimulates recruitment of eosinophils from the blood to sites of allergic inflammation. Mobilization from the bone marrow is an important early step in eosinophil trafficking during the allergic inflammatory response. In this paper we examine the potential of eotaxin to mobilize eosinophils and their progenitors from bone marrow. Eotaxin stimulated selective, dose-dependent chemotaxis of guinea pig bone marrow eosinophils in vitro. Intravenous injection of eotaxin (1 nmol/kg) into guinea pigs in vivo stimulated a rapid blood eosinophilia (from 3.9 ± 1.2 to 28 ± 9.9 × 104eosinophils/mL at 30 minutes) and a corresponding decrease in the number of eosinophils retained in the femoral marrow (from 9.0 ± 0.8 to 4.8 ± 0.8 × 106 eosinophils per femur). To show a direct release of eosinophils from the bone marrow an in situ perfusion system of the guinea pig femoral bone marrow was developed. Infusion of eotaxin into the arterial supply of the perfused femoral marrow stimulated a rapid and selective release of eosinophils into the draining vein. In addition, eotaxin stimulated the release of colony-forming progenitor cells. The cytokine interleukin-5 was chemokinetic for bone marrow eosinophils and exhibited a marked synergism with eotaxin with respect to mobilization of mature eosinophils from the femoral marrow. Thus, eotaxin may be involved in both the mobilization of eosinophils and their progenitors from the bone marrow into the blood and in their subsequent recruitment into sites of allergic inflammation.

scholarly journals Eotaxin Induces a Rapid Release of Eosinophils and Their Progenitors From the Bone Marrow

Blood ◽  
1998 ◽  
Vol 91 (7) ◽  
pp. 2240-2248 ◽  
Author(s):  
Roger T. Palframan ◽  
Paul D. Collins ◽  
Timothy J. Williams ◽  
Sara M. Rankin
Keyword(s):  
Bone Marrow ◽  
Guinea Pig ◽  
Allergic Inflammation ◽  
Interleukin 5 ◽  
In Situ Perfusion ◽  
Dose Dependent ◽  
Femoral Bone Marrow

Abstract The CC-chemokine eotaxin is a potent eosinophil chemoattractant that stimulates recruitment of eosinophils from the blood to sites of allergic inflammation. Mobilization from the bone marrow is an important early step in eosinophil trafficking during the allergic inflammatory response. In this paper we examine the potential of eotaxin to mobilize eosinophils and their progenitors from bone marrow. Eotaxin stimulated selective, dose-dependent chemotaxis of guinea pig bone marrow eosinophils in vitro. Intravenous injection of eotaxin (1 nmol/kg) into guinea pigs in vivo stimulated a rapid blood eosinophilia (from 3.9 ± 1.2 to 28 ± 9.9 × 104eosinophils/mL at 30 minutes) and a corresponding decrease in the number of eosinophils retained in the femoral marrow (from 9.0 ± 0.8 to 4.8 ± 0.8 × 106 eosinophils per femur). To show a direct release of eosinophils from the bone marrow an in situ perfusion system of the guinea pig femoral bone marrow was developed. Infusion of eotaxin into the arterial supply of the perfused femoral marrow stimulated a rapid and selective release of eosinophils into the draining vein. In addition, eotaxin stimulated the release of colony-forming progenitor cells. The cytokine interleukin-5 was chemokinetic for bone marrow eosinophils and exhibited a marked synergism with eotaxin with respect to mobilization of mature eosinophils from the femoral marrow. Thus, eotaxin may be involved in both the mobilization of eosinophils and their progenitors from the bone marrow into the blood and in their subsequent recruitment into sites of allergic inflammation.

scholarly journals Mechanisms of Acute Eosinophil Mobilization from the Bone Marrow Stimulated by Interleukin 5: The Role of Specific Adhesion Molecules and Phosphatidylinositol 3-Kinase

1998 ◽  
Vol 188 (9) ◽  
pp. 1621-1632 ◽  
Author(s):  
Roger T. Palframan ◽  
Paul D. Collins ◽  
Nicholas J. Severs ◽  
Stephen Rothery ◽  
Timothy J. Williams ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Guinea Pig ◽  
Light And Electron Microscopy ◽  
Phosphatidylinositol 3 Kinase ◽  
Release Process ◽  
Blocking Antibody ◽  
Β2 Integrin ◽  
Phosphatidylinositol 3

Mobilization of bone marrow eosinophils is a critical early step in their trafficking to the lung during allergic inflammatory reactions. We have shown previously that the cytokine interleukin (IL)-5, generated during an allergic inflammatory reaction in the guinea pig, acts systemically to mobilize eosinophils from the bone marrow. Here, we have investigated the mechanisms underlying this release process. Examination by light and electron microscopy revealed the rapid migration of eosinophils from the hematopoietic compartment and across the bone marrow sinus endothelium in response to IL-5. Using an in situ perfusion system of the guinea pig hind limb, we showed that IL-5 stimulated a dose-dependent selective release of eosinophils from the bone marrow. Eosinophils released from the bone marrow in response to IL-5 expressed increased levels of β2 integrin and a decrease in L-selectin, but no change in α4 integrin levels. A β2 integrin–blocking antibody markedly inhibited the mobilization of eosinophils from the bone marrow stimulated by IL-5. In contrast, an α4 integrin blocking antibody increased the rate of eosinophil mobilization induced by IL-5. In vitro we demonstrated that IL-5 stimulates the selective chemokinesis of bone marrow eosinophils, a process markedly inhibited by two structurally distinct inhibitors of phosphatidylinositol 3-kinase, wortmannin and LY294002. Wortmannin was also shown to block eosinophil release induced by IL-5 in the perfused bone marrow system. The parallel observations on the bone marrow eosinophil release process and responses in isolated eosinophils in vitro suggest that eosinophil chemokinesis is the driving force for release in vivo and that this release process is regulated by α4 and β2 integrins acting in opposite directions.

STUDIES ON IN VITRO ANTIBODY PRODUCTION: III. PRODUCTION OF COMPLEMENT

1965 ◽  
Vol 11 (3) ◽  
pp. 415-425 ◽  
Author(s):  
R. Siboo ◽  
S. I. Vas
Keyword(s):  
Bone Marrow ◽  
Guinea Pig ◽  
Hemolytic Activity ◽  
Bone Marrow Cells ◽  
Heat Inactivation ◽  
Liver Slices ◽  
C Proteins ◽  
Antigen Antibody ◽  
Thymus Cells

Bone marrow, spleen, liver, and thymus cells of the guinea pig were incubated in Eagle"s minimal essential medium which contained glycine-2-C14. Flasks were removed from the water bath at 0, 6, 18, 24, and 48 hours and their contents were homogenized, centrifuged, and concentrated. Experiments were carried out with the concentrated supernatant samples to determine whether or not they contained proteins with properties similar to those of complement (C′) proteins. Titrations for hemolytic activity revealed that bone marrow cells produced C′2, C′3, and C′4; spleen cells synthesized C′2 and liver slices produced C′2. Liver supernatant samples also contained C′3 activity, but proteins associated with C′3 actiyity seemed to be released rather than synthesized by liver slices. We were unable to demonstrate C activity with thymus cell supernatants. Heat inactivation (56 °C for 30 minutes), as well as EDTA inhibited the hemolytic activity of supernatant samples.Further experiments showed that these supernatant proteins were fixed to antigen–antibody complexes and to aggregated gamma-globulin. The fixation was inhibited by guinea pig serum. Immunoelectrophoresis and gel-diffusion showed that proteins synthesized in vitro migrated as beta-globulins and were antigenically similar to C′ proteins.

In vitro derivation of macrophage from guinea pig bone marrow with human M-CSF

2013 ◽  
Vol 389 (1-2) ◽  
pp. 88-94
Author(s):  
Karl O.A. Yu ◽  
Steven A. Porcelli ◽  
Howard A. Shuman
Keyword(s):  
Bone Marrow ◽  
Guinea Pig

Thrombocytopenia in Guinea Pigs Infected by Encephalomyocarditis Virus (EMC)

1967 ◽  
Vol 18 (03/04) ◽  
pp. 686-690
Author(s):  
Y Modai ◽  
R Oren ◽  
A de Vries ◽  
A Kohn
Keyword(s):  
Bone Marrow ◽  
Guinea Pig ◽  
Guinea Pigs ◽  
Encephalomyocarditis Virus ◽  
Bone Marrow Culture ◽  
Emc Virus ◽  
Bone Marrow Cultures ◽  
Intraperitoneal Infection ◽  
Marrow Cultures

SummaryIntraperitoneal infection of guinea pigs with Encephalomyocarditis (EMC) virus led to viremia 2-10 days after infection and to paralysis and death of some of the infected animals. During the course of infection there was marked leucocytosis and thrombocytopenia. No abnormalities were detected in megakaryocytes in bone marrow cultures from infected guinea pigs. Exposure of guinea pig bone marrow culture to EMC virus in vitro impaired granulation and disintegration of megakaryocytes to platelets.

scholarly journals The Effect of Erythrocytes on Mouse Bone Marrow Colony Development In Vitro

Blood ◽  
1971 ◽  
Vol 38 (3) ◽  
pp. 353-359 ◽  
Author(s):  
T. R. BRADLEY ◽  
P. A. TELFER ◽  
PHYLLIS FRY
Keyword(s):  
Bone Marrow ◽  
Guinea Pig ◽  
Culture System ◽  
Red Cells ◽  
Colony Development ◽  
Mouse Bone Marrow ◽  
Maximal Dose ◽  
Colony Forming Units ◽  
Development In Vitro

Abstract The development of colonies in nutrient agar in vitro from mouse hemopoietic cells, in the presence of a maximal dose of colony-stimulating activity (CSA), was greatly enhanced by the addition of washed red cells to the culture system. Human, sheep, guinea pig, rat, and mouse (syngeneic or allogeneic), but not rabbit, red cells exhibited enhancing ability, as did osmotically lysed erythrocyte preparations from rats. The use of red cells in the culture system, in addition to a maximum concentration of CSA, has resulted in revised estimates of the numbers of agar colony-forming units in hemopoietic tissues.

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