Erythroid precursors studied by mouse bone marrow cultivation in a plasma clot]

Two types of erythroid precursors were found by cultivation of the mouse bone marrow in the plasma clot with mouse serum and without adding exogenic erythropoietin to the culture medium. The first precursor had properties similar to the erythroid colony-forming unit (CFUe) previously described while the second resembles in its properties the erythroid burst-forming unit (BFUe). Optimal concentration of mouse serum in the culture medium was 10-15%. Clone nature of the colonies and bursts described is confirmed by linear dependence of their number on the cell concentration in the culture.     

Immunoglobulin-positive cells in mouse bone marrow]

The content of Ig-bearing lymphocytes and their precursors in the mouse bone marrow was investigated 6 and 36 hours after the hydroxyurea treatment. Some increase of the B-cell content takes place in the trated bone marrow. Dividing and non-dividing B-cell precursors, except the stem cells, were practically absent.     

Identifying superoxide-sensitive progenitor cells in mouse bone marrow

Macrophage progenitor cells in bone marrow, that develop into attached colonies in liquid culture medium, contain a fraction of cells sensitive to photochemically generated superoxide radicals. This fraction varies from one animal to another. Populations of cells containing the superoxide-sensitive fraction show a greater sensitivity to X-rays than do populations in which this fraction has been photochemically inactivated. The change in radiosensitivity was proportional to the superoxide-sensitive fraction.     

Histamine production by alloantigen-activated mouse bone marrow cells

Histamine's contribution to the manifestations associated with graft-versus-host disease (GVHD) and/or hybrid resistance is unknown. Thus, we initiated studies to see whether or not mouse bone marrow cells could produce histamine upon alloantigen stimulation. Irradiated allogeneic spleen cells were shown to stimulate bone marrow cells to produce and secrete high levels of histamine. During 7 days of culture there was only a marginal increase in cell-associated histamine while the amount of histamine in the supernatant increased 10- to 20-fold. Optimal histamine production was dependent upon Lyt 1+2+ T cells resident in the bone marrow. Further, bone marrow cells from Nude mice failed to produce high levels of histamine following alloantigen stimulation. Soluble factors produced by alloantigen-stimulated bone marrow cells or by Con A-stimulated rat spleen cells induced high levels of histamine production in bone marrow cells in the absence of alloantigen. We suggest that histamine production by alloantigen-activated bone marrow cells may modulate immune functions following bone marrow transplantation.     

Clastogenic effects of acrylamide in mouse bone marrow cells

Acrylamide, known to induce dominant-lethal mutations (Shelby et al., 1986; Smith et al., 1986) and heritable translocations (Shelby et al., 1987) in rodent germ cells, was hitherto a questionable clastogen in rodent bone marrow (Shiraishi, 1978). Therefore, it was tested for chromosomal aberrations in mouse bone marrow cells, spermatogonia and by the micronucleus test. The intraperitoneally injected doses ranged from 50 to 150 mg/kg. In the chromosomal bone marrow test and the micronucleus assay positive results were obtained with acrylamide, and in the latter test the effect increased linearly with dose. Chromosomal aberrations were not induced in differentiating spermatogonia by the acute acrylamide treatment. Cisplatin was used as a positive control and gave the expected positive response in all 3 tests. The present results demonstrate that acrylamide is no exception among clastogens. It breaks chromosomes not only in mammalian germ cells but also in somatic cells.     

Clastogenicity of carbazole in mouse bone marrow cells in vivo

The protective effect of Nigella sativa seed extract and its main constituents thymoquinone (TQ) was studied on mouse cells infected with schistosomiasis. Bone marrow cells in the in vivo experiments and spleen cells in the in vitro one were used to evaluate the potentially protective effect of these natural compounds on the induction of chromosomal aberrations. Karyotyping of the mice cells illustrated that the main abnormalities were gaps, fragments and deletions especially in chromosomes 2, 6 and some in chromosomes 13 and 14. Both N. sativa extract and TQ were considered as protective agents against the chromosomal aberrations induced as a result of schistosomiasis.     

Clastogenicity of carbazole in mouse bone marrow cells in vivo

Clastogenicity of carbazole was evaluated by employing mouse in vivo chromosomal aberration (CA) test. Carbazole administered intraperitoneally (i.p.) at the rate of 25, 50, 100, 150 and 200 mg/kg b.w. to Swiss albino mice in vivo resulted in mitotic depression and induction of chromosomal aberrations. Dose related decrease in mitotic index (MI) and increase in the frequencies of chromosomal aberrations per cell (CAs/cell) and percent abnormal cells were recorded in bone marrow cells. However, statistically significant reduction in MI and increase in CAs/cell and percent abnormal cells were found only for the two higher doses. The results obtained indicate that carbazole or its metabolite, if any, is moderately clastogenic in the bone marrow cells of Swiss albino mice.     

Cultivation of frozen mouse bone marrow cells in agar.

The authors attempted to cultivate frozen mouse bone marrow cells in a semisolid medium. They demonstrated that the stem haematopoietic cells of frozen mouse bone marrow were capable of proliferation and of colony formation on agar. The much smaller number of colonies from frozen mouse bone marrow (about 80% fewer) compared with fresh marrow is evidence that part of the stem haematopoietic cell population retains proliferative capacity even after freezing.     

Aberrant immunoglobulin gene rearrangement in scid mouse bone marrow cells

We have analyzed Ig gene rearrangement in the immunodeficient mutant mouse, CB-17 scid. Bone marrow stem cells from scid mice were cultured in the in vitro culture system of Whitlock and Witte. Ig gene rearrangement in the scid cells was studied by DNA cloning. Seven DNA clones of Ig H chain JH and DH regions were analyzed by DNA sequencing, and all the clones contained a failure in D-J joining. In the rearranged structure, both DH and JH coding sequences are either partly or completely deleted. Molecular mechanisms causing the aberrant DNA rearrangement are discussed.     

Osteogenic differentiation of mouse bone marrow stromal cells on fibroin microcarriers

We investigated the proliferation and osteogenic differentiation of mesenchymal stem cells cultured on fibroin microcarriers. Effective cell proliferation on the surface of the microcarriers, determined by the large surface area, and the contribution of microcarrier mineralization to the stimulation of the osteogenic differentiation of mesenchymal stem cells was revealed.     

A method for generation of bone marrow-derived macrophages from cryopreserved mouse bone marrow cells

The broad use of transgenic and gene-targeted mice has established bone marrow-derived macrophages (BMDM) as important mammalian host cells for investigation of the macrophages biology. Over the last decade, extensive research has been done to determine how to freeze and store viable hematopoietic human cells; however, there is no information regarding generation of BMDM from frozen murine bone marrow (BM) cells. Here, we establish a highly efficient protocol to freeze murine BM cells and further generate BMDM. Cryopreserved murine BM cells maintain their potential for BMDM differentiation for more than 6 years. We compared BMDM obtained from fresh and frozen BM cells and found that both are similarly able to trigger the expression of CD80 and CD86 in response to LPS or infection with the intracellular bacteria Legionella pneumophila. Additionally, BMDM obtained from fresh or frozen BM cells equally restrict or support the intracellular multiplication of pathogens such as L. pneumophila and the protozoan parasite Leishmania (L.) amazonensis. Although further investigation are required to support the use of the method for generation of dendritic cells, preliminary experiments indicate that bone marrow-derived dendritic cells can also be generated from cryopreserved BM cells. Overall, the method described and validated herein represents a technical advance as it allows ready and easy generation of BMDM from a stock of frozen BM cells.     

Effect of plutonium-239 on the mitotic activity of mouse bone marrow cells

Summary Mitotic index of the bone marrow cells was studied in femoral bone marrow of mice given 313 kBq²³⁹Pu kg^–1. The attention was turned to the femoral midshaft and the mitose concentration, intensified by Colcemid stathmokinetic effect, was evaluated in a sampling field from endosteal surface to the central venous canal, throughout 68 weeks. It has been found that the plutonium effect in the sampling band is rather uniform except the points in subendosteal zone early after plutonium injection, where the mitotic index was reduced in such a way that the mitotic gradient, observed in controls, was affected. The mitotic activity in femoral diaphysis of plutonium injected mice was mobilized approximately till the 30th week of contamination. Later it deteriorated progressively. The results are discussed and should not be regarded as representative for the entire bone marrow hemopoiesis.     

Osteogenic properties of adhesive cells in Dexter culture of the mouse bone marrow

Disaggregated cell suspensions obtained by mouse bone marrow fermentative digestion as well as stromal tissue obtained by marrow mild mechanical destruction were explanted. Both methods yield the cultures in which the hematopoiesis duration is comparable with dexter cultures. Adhesive cells from all of these three culture types were resuspended and in the porous gelatin sponges heterotopically transplanted under the kidney capsule of syngenic recipients. In the transplantation site there develops the hemopoietic organ containing reticular stroma, hemopoietic cells, and in most cases the well developed bone tissue. Thus, the adherent layers of mouse bone marrow dexter and similar cultures contain for a long period (not less than 2-3.5 months) the stromal fibroblast population which maintains its osteogenic and hemopoietic microenvironment transfer capacities.