scholarly journals Assessment of erythrocytic and granulocytic colony formation in an in vivo plasma clot diffusion chamber culture system

Blood ◽  
1976 ◽  
Vol 47 (6) ◽  
pp. 1041-1051
Author(s):  
HN Steinberg ◽  
ES Handler ◽  
EE Handler
Keyword(s):  
Colony Formation ◽  
Bone Marrow Cells ◽  
Secondary Growth ◽  
Diffusion Chamber ◽  
Plasma Clot ◽  
Granulocytic Colony ◽  
Normal Rat ◽  
Diffusion Chamber Culture ◽  
Marrow Cells

Normal rat bone marrow cells seeded into a plasma clot diffusion chamber culture developed into erythrocytic and granulocytic colonies in vivo. Chambers implanted into the peritoneal cavity of normal hosts showed erythrocytic colony numbers reaching an initial peak on day 2, declining on days 3--5, and increasing in a secondary growth phase on day 7. Day 2 colonies were evenly dispersed; day 7 colonies were grouped into discrete areas of bursts. Granulocytic colony numbers reached a peak on day 4 and gradually declined through day 7. Cells in various stages of differentiation could be detected in both colony types. Colony numbers were proportional to the number of marrow cells seeded into the chamber. Host animals treated with phenylhydrazine induced a marked increase in erythrocytic colony numbers and size and a decrease in granulocytic colony formation. Host animals treated with endotoxin suppressed erythrocytic colonies while increasing granulocytic colony size. This method may prove advantageous for the study of hematopoietic colony formation in a physiologic environment.

scholarly journals Assessment of erythrocytic and granulocytic colony formation in an in vivo plasma clot diffusion chamber culture system

Blood ◽  
1976 ◽  
Vol 47 (6) ◽  
pp. 1041-1051 ◽  
Author(s):  
HN Steinberg ◽  
ES Handler ◽  
EE Handler
Keyword(s):  
Colony Formation ◽  
Bone Marrow Cells ◽  
Secondary Growth ◽  
Diffusion Chamber ◽  
Plasma Clot ◽  
Granulocytic Colony ◽  
Normal Rat ◽  
Diffusion Chamber Culture ◽  
Marrow Cells

Abstract Normal rat bone marrow cells seeded into a plasma clot diffusion chamber culture developed into erythrocytic and granulocytic colonies in vivo. Chambers implanted into the peritoneal cavity of normal hosts showed erythrocytic colony numbers reaching an initial peak on day 2, declining on days 3--5, and increasing in a secondary growth phase on day 7. Day 2 colonies were evenly dispersed; day 7 colonies were grouped into discrete areas of bursts. Granulocytic colony numbers reached a peak on day 4 and gradually declined through day 7. Cells in various stages of differentiation could be detected in both colony types. Colony numbers were proportional to the number of marrow cells seeded into the chamber. Host animals treated with phenylhydrazine induced a marked increase in erythrocytic colony numbers and size and a decrease in granulocytic colony formation. Host animals treated with endotoxin suppressed erythrocytic colonies while increasing granulocytic colony size. This method may prove advantageous for the study of hematopoietic colony formation in a physiologic environment.

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

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 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 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

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 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 CD34+ human marrow cells that express low levels of Kit protein are enriched for long-term marrow-engrafting cells

Blood ◽  
1996 ◽  
Vol 87 (10) ◽  
pp. 4136-4142 ◽  
Author(s):  
I Kawashima ◽  
ED Zanjani ◽  
G Almaida-Porada ◽  
AW Flake ◽  
H Zeng ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Marrow Cell ◽  
In Utero ◽  
Fetal Sheep ◽  
Hematopoietic Stem ◽  
Show Evidence ◽  
Marrow Cells

Using in utero transplantation into fetal sheep, we examined the capability of human bone marrow CD34+ cells fractionated based on Kit protein expression to provide long-term in vivo engraftment. Twelve hundred to 5,000 CD34+ Kit-, CD34+ Kit(low), and CD34+ Kit(high) cells were injected into a total of 14 preimmune fetal sheep recipients using the amniotic bubble technique. Six fetuses were killed in utero 1.5 months after bone marrow cell transplantation. Two fetuses receiving CD34+ Kit(low) cells showed signs of engraftment according to analysis of CD45+ cells in their bone marrow cells and karyotype studies of the colonies grown in methylcellulose culture. In contrast, two fetuses receiving CD34+ Kit(high) cells and two fetuses receiving CD34+ Kit- cells failed to show evidence of significant engraftment. Two fetuses were absorbed. A total of six fetuses receiving different cell populations were allowed to proceed to term, and the newborn sheep were serially examined for the presence of chimerism. Again, only the two sheep receiving CD34+ Kit(low) cells exhibited signs of engraftment upon serial examination. Earlier in studies of murine hematopoiesis, we have shown stage-specific changes in Kit expression by the progenitors. The studies of human cells reported here are in agreement with observations in mice, and indicate that human hematopoietic stem cells are enriched in the Kit(low) population.

Distribution of sister chromatid exchanges on the mouse chromosomes in vivo with reference to the replication properties of the X chromosome

1984 ◽  
Vol 26 (2) ◽  
pp. 152-157
Author(s):  
S. M. Singh ◽  
D. L. Reimer
Keyword(s):  
Bone Marrow ◽  
X Chromosome ◽  
Sister Chromatid ◽  
Bone Marrow Cells ◽  
Relative Length ◽  
Chromosome Length ◽  
Sister Chromatid Exchanges ◽  
Chromatid Exchanges ◽  
Marrow Cells

Frequency of sister chromatid exchanges (SCE) were recorded separately for different chromosomes from bone marrow cells of female mice of the two genetic strains (C3H/S and C57BL/6J). SCEs were evaluated following different doses of 5-bromo-2′deoxyuridine (BrdU) as nine hourly i.p. injections. The SCE per cell increased with increasing BrdU doses which was slightly higher in C3H/S than in the C57BL/6J. SCEs per cell were variable at every treatment – strain combination, possibly reflecting the heterogeneous nature of the bone marrow cells. In general, there is a positive correlation between SCE per chromosome and the relative chromosome length. Total SCEs on one of the large chromosomes (most likely the X chromosome), however, are significantly higher than expected on the basis of relative length alone. Most of this increase is attributable to one of the homologues of this chromosome, which is not in synchrony with the rest of the chromosomes and may represent the late-replicating X. These results when viewed in the light of replication properties of the heterochromatinized X, suggest a direct involvement of DNA replication in SCE formation and may argue against the replication point as the sole site for the SCEs.Key words: sister chromatid exchange, BrdU, recombination, replication, X chromosome.

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