scholarly journals Radioautographic Studies of Bone Marrow Lymphocytes in Vivo and in Diffusion Chamber Cultures

Blood ◽  
1964 ◽  
Vol 23 (1) ◽  
pp. 1-17 ◽  
Author(s):  
D. G. OSMOND ◽  
N. B. EVERETT
Keyword(s):  
Bone Marrow ◽  
Large Scale ◽  
Bone Marrow Cells ◽  
Tritiated Thymidine ◽  
Diffusion Chamber ◽  
Blood Stream ◽  
Turnover Time ◽  
Diffusion Chambers ◽  
Transitional Cells

Abstract Radioautography with tritiated thymidine has been utilized to examine the turnover rate and origin of small lymphocytes in the bone marrow of the guinea-pig. Very few marrow lymphocytes were initially labeled by a single injection of tritiated thymidine, but thereafter the number of labeled lymphocytes rapidly increased to high maximum levels at 3 days. Analysis of the labeling curves and grain counts indicates that the population of marrow lymphocytes is maintained in a dynamic steady state with an average turnover time of 3 days or less. Suspensions of bone marrow cells were isolated from the circulation within intraperitoneal diffusion chambers after short-term labeling with tritiated thymidine in vivo. Although very few small lymphocytes were labeled when introduced into the diffusion chambers, a considerable percentage became labeled during the subsequent culture period. Tritiated thymidine was also administered intravenously whilst excluded from one hind limb by the application of an occlusive compression bandage for 20 minutes. Very few labeled small lymphocytes were found after 72 hours in the tibial marrow of the initially occluded limb, whereas the normal high percentage was labeled in the control tibial marrow. These experiments do not demonstrate any large-scale influx of small lymphocytes from the blood stream into the marrow parenchyma. They suggest that newly formed small lymphocytes appear in the marrow as a result of the division of locally situated precursor cells, but the mechanism of intramedullary lymphocytopoiesis is uncertain. "Transitional" cells, intermediate in morphology between blast cells and small lymphocytes, synthesize DNA and are actively proliferative, but they do not appear to account fully for the rate of lymphocyte production. Certain large, undifferentiated labeled cells appeared in the bone marrow as a result of hematogenous migration. Some implications of these findings are discussed.

scholarly journals Further evidence of the in vivo role of erythropoietin or companion molecules induced by hypoxia on proliferation and continuing differentiation of BFU-e in PCDC

Blood ◽  
1981 ◽  
Vol 57 (2) ◽  
pp. 298-304
Author(s):  
K Harigaya ◽  
EP Cronkite ◽  
ME Miller ◽  
G Moccia
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Diffusion Chamber ◽  
Hypoxic Exposure ◽  
Red Cell ◽  
Plasma Clot ◽  
Diffusion Chambers ◽  
Cell Production

Normal and plethoric bone marrow cells were grown in plasma clot diffusion chambers (PCDC) implanted into the peritoneum of normal mice or mice submitted to 7 her of hypoxia (23,000 ft) daily, on a single day or on 2 consecutive days at different times after implantation of the PCDC's. Daily discontinuous hypoxia (DDH) produced more 6-day bursts than other treatments. Hypoxia on days 1 and 2 after implantation was nearly as effective as DDH on day-6 bursts. Later bouts of hypoxia or a singly hypoxic exposure on day 1 or 2 was less effective. Erythropoietin (Ep) levels were measured by bioassay on both diffusion chamber (DC) contents and serum. Serum Ep levels peaked at 160 mU/ml after a 7-hr hypoxic exposure while the DC content Ep levels were in the nondetectable range (less than 50 mU/ml). The data implies that either higher than normal Ep levels or a companion molecules (s) produced by hypoxia are required for 1–2 days early in the culture period of force an increasing number of BFU-d-e down the erythrocytic pathway and thus increase red cell production at times of need in vivo.

scholarly journals Potentiality of the murine colony-forming cells detected by an in vivo diffusion chamber culture system

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 686-689
Author(s):  
E Niskanen ◽  
HE Wyandt
Keyword(s):  
Bone Marrow ◽  
Colony Formation ◽  
Culture System ◽  
Bone Marrow Cells ◽  
Diffusion Chamber ◽  
Diffusion Chambers ◽  
Hemopoietic Cells ◽  
Chromosome Marker ◽  
Diffusion Chamber Culture

Culture of a mixture of bone marrow cells with and without T6 chromosome marker in diffusion chambers in mice yielded colonies (CFU- DG) containing cells of a single karyotype, suggesting clonality. Injection of individual CFU-DG colonies into lethally irradiated mice resulted in increased spleen colony formation on day 12 (CFU-S). The possibility of endogenous origin was excluded by demonstrating the presence of T6 marker in both CFU-DG and CFU-S colonies in karyotypically normal hosts. These findings suggest that the cells giving rise to granulocytic colonies in diffusion chambers also can give rise to multipotential hemopoietic cells.

scholarly journals Potentiality of the murine colony-forming cells detected by an in vivo diffusion chamber culture system

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 686-689 ◽  
Author(s):  
E Niskanen ◽  
HE Wyandt
Keyword(s):  
Bone Marrow ◽  
Colony Formation ◽  
Culture System ◽  
Bone Marrow Cells ◽  
Diffusion Chamber ◽  
Diffusion Chambers ◽  
Hemopoietic Cells ◽  
Chromosome Marker ◽  
Diffusion Chamber Culture

Abstract Culture of a mixture of bone marrow cells with and without T6 chromosome marker in diffusion chambers in mice yielded colonies (CFU- DG) containing cells of a single karyotype, suggesting clonality. Injection of individual CFU-DG colonies into lethally irradiated mice resulted in increased spleen colony formation on day 12 (CFU-S). The possibility of endogenous origin was excluded by demonstrating the presence of T6 marker in both CFU-DG and CFU-S colonies in karyotypically normal hosts. These findings suggest that the cells giving rise to granulocytic colonies in diffusion chambers also can give rise to multipotential hemopoietic cells.

scholarly journals Further evidence of the in vivo role of erythropoietin or companion molecules induced by hypoxia on proliferation and continuing differentiation of BFU-e in PCDC

Blood ◽  
1981 ◽  
Vol 57 (2) ◽  
pp. 298-304 ◽  
Author(s):  
K Harigaya ◽  
EP Cronkite ◽  
ME Miller ◽  
G Moccia
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Diffusion Chamber ◽  
Hypoxic Exposure ◽  
Red Cell ◽  
Plasma Clot ◽  
Diffusion Chambers ◽  
Cell Production

Abstract Normal and plethoric bone marrow cells were grown in plasma clot diffusion chambers (PCDC) implanted into the peritoneum of normal mice or mice submitted to 7 her of hypoxia (23,000 ft) daily, on a single day or on 2 consecutive days at different times after implantation of the PCDC's. Daily discontinuous hypoxia (DDH) produced more 6-day bursts than other treatments. Hypoxia on days 1 and 2 after implantation was nearly as effective as DDH on day-6 bursts. Later bouts of hypoxia or a singly hypoxic exposure on day 1 or 2 was less effective. Erythropoietin (Ep) levels were measured by bioassay on both diffusion chamber (DC) contents and serum. Serum Ep levels peaked at 160 mU/ml after a 7-hr hypoxic exposure while the DC content Ep levels were in the nondetectable range (less than 50 mU/ml). The data implies that either higher than normal Ep levels or a companion molecules (s) produced by hypoxia are required for 1–2 days early in the culture period of force an increasing number of BFU-d-e down the erythrocytic pathway and thus increase red cell production at times of need in vivo.

scholarly journals Proliferation of diffusion chamber colony-forming units (CFUD) in cultures of normal human bone marrow in diffusion chambers in mice

Blood ◽  
1977 ◽  
Vol 49 (3) ◽  
pp. 415-424
Author(s):  
N Jacobsen
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Diffusion Chambers ◽  
Colony Forming Units ◽  
Normal Human ◽  
Normal Human Bone Marrow

Normal human bone marrow contains cells capable of forming colonies of hemopoietic cells in fibrin clots in diffusion chambers implanted intraperitoneally (i.p.) into irradiated mice. The present paper describes the proliferation of such colony-forming units (CFUD) in cultures in vivo. Cells harvested from diffusion chambers after 1–14 days of culture in 450-R irradiated mice contained CFUD, which formed neutrophilic, eosinophilic, or megakaryocytic colonies when tested by secondary culture in fibrin clot chambers. When bone marrow was precultured in irradiated mice at a concentration of 10(6) cells per chamber, an initial fall in the number of neutrophilic CFUD was observed. This decrease was followed by an increase to a maximum at day 2, and then a secondary decrease. The number of neutrophilic CFUD recovered after 2 days of preculture in irradiated mice varied between 60% and 250% of the number present before preculture. Preculture in nonirradiated mice resulted in a significantly lower recovery of neutrophilic CFUD. In vitro treatment of bone marrow cells with hydroxyurea (OHU) after 2 days of preculture in irradiated mice resulted in a 68% +/- 5% reduction in the number of neutrophilic CFUD. In contrast, OHU had no similar effect on precultures from nonirradiated mice. Both the recovery and sensitivity to OHU of eosinophilic CFUD were independent of host irradiation. Similarly, no effect of host irradiation on the recovery or the 3H-thymidine (3HTdR) labeling index of morphologically recognizable granulocytic cells was observed at day 2. The data suggest an effect of humoral host factor(s) on the proliferation of early precursor cells, which are or become committed to differentiate into the neutrophilic pathway in diffusion chambers.

scholarly journals The Cultivation of Mouse Bone Marrow in Vivo

Blood ◽  
1959 ◽  
Vol 14 (9) ◽  
pp. 1040-1046 ◽  
Author(s):  
IRWIN BERMAN ◽  
HENRY S. KAPLAN
Keyword(s):  
Bone Marrow ◽  
Peritoneal Cavity ◽  
Normal Mouse ◽  
Bone Marrow Cells ◽  
Mouse Bone Marrow ◽  
Considerable Time ◽  
Diffusion Chambers ◽  
Mouse Bone Marrow Cells ◽  
Marrow Cells

Abstract The cultivation of normal mouse bone marrow cells in diffusion chambers implanted into the peritoneal cavity of mice has been described. Mouse bone marrow cells cultivated by this method continue to undergo differentiation and maintain their morphologic identity for a considerable time.

Stimulation of monocyte precursors in vivo by an extract from Listeria monocytogenes

1979 ◽  
Vol 25 (6) ◽  
pp. 698-705 ◽  
Author(s):  
D. T. Shum ◽  
S. B. Galsworthy
Keyword(s):  
Bone Marrow ◽  
Listeria Monocytogenes ◽  
Generation Time ◽  
Bone Marrow Cells ◽  
Tritiated Thymidine ◽  
Water Soluble ◽  
Blood Monocytes ◽  
Expected Time ◽  
Stimulation Of

A water-soluble monocytosis-producing activity (MPA) extracted from Listeria monocytogenes was found to stimulate proliferation of promonocytes in vivo. Mice were pulse-labelled for 2 h with tritiated thymidine ([3H]TdR) at various times after intraperitoneal injection of MPA. Autoradiography of bone marrow cells revealed an increased labelling index of promonocytes of MPA-treated mice which was maximum 8 h after the MPA injection. Mice labelled with [3H]TdR 8 h after MPA injection developed a monocytosis at the expected time (peak at 48 h) and the blood monocytes were found to be highly labelled. Both the generation time of monocyte precursors and the halftime of blood monocytes were found to be shorter than the corresponding values in control mice.

scholarly journals Human T-lymphocyte products stimulate human hemopoietic progenitor cell proliferation in diffusion chambers in vivo

Blood ◽  
1982 ◽  
Vol 60 (2) ◽  
pp. 368-372 ◽  
Author(s):  
E Niskanen ◽  
A Oki ◽  
MJ Cline ◽  
DW Golde
Keyword(s):  
Cell Line ◽  
Conditioned Medium ◽  
Colony Formation ◽  
Bone Marrow Cells ◽  
Diffusion Chamber ◽  
Heat Inactivation ◽  
Diffusion Chambers ◽  
Human T Cell

Abstract Human myeloid colony formation in diffusion chambers in mice (CFU-DG) was enhanced following administration of a human T-cell-line-derived conditioned medium (Mo). The Mo cell line also elaborates activities stimulating human myeloid colony formation in vitro in agar (CSF) and potentiating erythroid colony formation in vitro in methylcellulose (EPA). Depletion of CSF from Mo conditioned medium by heat inactivation or gel exclusion chromatography did not affect CFU-DG formation. EPA and CFU-DG stimulating activities are heat stable and have approximately the same molecular weight. Culture of human bone marrow cells in diffusion chambers in mice for 4 days under the influence of Mo conditioned medium resulted in significant increment of BFU-E and CFU-DG as judged by subculture of diffusion chamber contents. No effect on CFU-C could be detected.

scholarly journals The relative spatial distributions of CFUs and CFUc in the normal mouse femur

Blood ◽  
1975 ◽  
Vol 46 (1) ◽  
pp. 65-72 ◽  
Author(s):  
BI Lord ◽  
NG Testa ◽  
JH Hendry
Keyword(s):  
Bone Marrow ◽  
Spatial Organization ◽  
Bone Marrow Cells ◽  
Tritiated Thymidine ◽  
Marrow Cell ◽  
Bone Surface ◽  
Femoral Bone Marrow ◽  
Peak Value ◽  
Marrow Cells

Femoral bone marrow was divided longitudinally into two groups of cells of varying size. By assaying CFU and CFU in the two zones of the marrow, their distributions across the diameter of the femur was determined. It is shown that the concentration of CFU increases from the femoral axis (15 CFU/105 bone marrow cells) to the bone surface (44 CFU/105 cells), obeying approximately a square-law relationship. The CFU concentration, on the other hand, increases from the femoral axis (32CFU/105 cells) to a peak value (260 CFU/105 cells) at about 330 um from the axis and thence falls off against to the bone surface (77 CFU/105 cells). Selective kinning cells in DNA synthesis using the tritiated thymidine suicide technique, in vivo, showed that CFU, near the bone surface are proliferating at a faster rate than those more distant from bone, but that CFU have a fast proliferation rate irrespective of their position in the distribution. Thus, bone marrow cell populations are shown to conform to a well-defined spatial organization corresponding to the chronologic relationships between marrow cells.

scholarly journals Proliferation of diffusion chamber colony-forming units (CFUD) in cultures of normal human bone marrow in diffusion chambers in mice

Blood ◽  
1977 ◽  
Vol 49 (3) ◽  
pp. 415-424 ◽  
Author(s):  
N Jacobsen
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Diffusion Chambers ◽  
Colony Forming Units ◽  
Normal Human ◽  
Normal Human Bone Marrow

Abstract Normal human bone marrow contains cells capable of forming colonies of hemopoietic cells in fibrin clots in diffusion chambers implanted intraperitoneally (i.p.) into irradiated mice. The present paper describes the proliferation of such colony-forming units (CFUD) in cultures in vivo. Cells harvested from diffusion chambers after 1–14 days of culture in 450-R irradiated mice contained CFUD, which formed neutrophilic, eosinophilic, or megakaryocytic colonies when tested by secondary culture in fibrin clot chambers. When bone marrow was precultured in irradiated mice at a concentration of 10(6) cells per chamber, an initial fall in the number of neutrophilic CFUD was observed. This decrease was followed by an increase to a maximum at day 2, and then a secondary decrease. The number of neutrophilic CFUD recovered after 2 days of preculture in irradiated mice varied between 60% and 250% of the number present before preculture. Preculture in nonirradiated mice resulted in a significantly lower recovery of neutrophilic CFUD. In vitro treatment of bone marrow cells with hydroxyurea (OHU) after 2 days of preculture in irradiated mice resulted in a 68% +/- 5% reduction in the number of neutrophilic CFUD. In contrast, OHU had no similar effect on precultures from nonirradiated mice. Both the recovery and sensitivity to OHU of eosinophilic CFUD were independent of host irradiation. Similarly, no effect of host irradiation on the recovery or the 3H-thymidine (3HTdR) labeling index of morphologically recognizable granulocytic cells was observed at day 2. The data suggest an effect of humoral host factor(s) on the proliferation of early precursor cells, which are or become committed to differentiate into the neutrophilic pathway in diffusion chambers.

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