Colony promoting activity and its target 'pre-CFUc' in genetically anemic mice of W/Wv genotype

Incubation of bone marrow cells with supernatant from long-term cultures of bone marrow cells increases the number of granulocyte-macrophage progenitor cells. This study reveals the presence of target cells of the colony promoting activity (CPA) in W/Wv mouse marrow. It is also shown that CPA does not stimulate erythroid colony formation in vitro.     

Granulocyte colony formation in vitro: enhancement by human placental (umbilical cord) serum.

Addition of human placental umbilical cord serum to bone marrow cultures reproducibly increased the number of granulocyte colonies in vitro. This stimulatory effect was significantly greater than that of fetal calf serum which was seen in cultures of human bone marrow under the conditions described.     

Macrophages derived from bone marrow modulate differentiation of myeloid dendritic cells

Dendritic cells (DC) are specialized antigen-presenting cells. Bone marrow monocytes have been widely used to generate murine myeloid DC. We found that mouse macrophages derived from bone marrow CD11b⁺ monocytes influenced the differentiation of these precursors into DC. Modulation of differentiation was demonstrated by the down-regulation of CD11c, CD40, and CD86 expression and by IL-12 production. DC differentiated in the presence of conditioned medium from bone marrow-derived macrophage culture (MCM) had impaired ability to stimulate proliferation of, and IFN- γ production by, allogeneic CD4⁺ T cells. This inhibition of DC differentiation was mainly mediated by secretory products from macrophages but not by cell-cell contact. MCM contained higher concentrations of macrophage-colony-stimulating factor (M-CSF), IL-10, and TGF- β1, whereas IL-6 remained unchanged compared with conditioned medium from fresh monocytes. M-CSF may be the major mediator in MCM inhibiting DC differentiation. This study demonstrates an important influence of bone marrow-derived macrophages on DC precursors during DC differentiation. Received 12 September 2006; received after revision 20 October 2006; accepted 13 November 2006     

The extracellular matrix of the hematopoietic microenvironment

The bone marrow microenvironment plays an important role in promoting hematopoietic progenitor cell proliferation and differentiation and the controlled egress of these developing hematopoietic cells. The establishment of long-term bone marrow cultures, which are thought to mimic hematopoiesis in vitro, and various stromal cell lines has greatly facilitated the analysis of the functions of this microenvironment. Extracellular matrix (ECM) molecules of all three categories (collagens, proteoglycans and glycoproteins) have been identified as part of this microenvironment and have been shown to be involved in, different biological functions such as cell adhesion and anti-adhesion, binding and presentation of various cytokines and regulation of cell growth. It is suggested that these matrix molecules in combination with cytokines are crucial for compartmentalization of the bone marrow. Although many cell adhesion molecules have been characterized on the surface of hematopoietic progenitor cells, the nature of cellular receptors for the ECM components is less well defined. During leukemia, many immature blood cells are released from bone marrow, but it is not yet known whether these abnormalities in hematopoiesis are also caused by an altered microenvironment or altered composition of its extracellular matrix. The elucidation of the involvement of specific ECM-isoforms and as yet not characterized ECM components and their receptors in the bone marrow will certainly help towards a better understanding of these phenomena.     

Effect of heat-labile enterotoxin (LT) produced byEscherichia coli on in vitro cell proliferation

Summary Filtrates fromE. coli H10407 cultures, giving a positive response for heat-labile enterotoxin (LT) in the Y-1 cell test, show an inhibitory activity both on³H-thymidine uptake by Ehrlich ascites cells and on granulocytic-macrophagic precursors (CFU-C) in murine bone marrow.Acknowledgments. We gratefully acknowledge the expert technical assistance of L. Basso and A. Gerosa of Department of Pathology, Hospital of Desio.     

In vitro cytostatic effect of adenine-arabinoside (Ara-A) and cytosine-arabinoside (Ara-C)

Summary Adenine-arabinoside, a new antiviral drug with questionable bone marrow toxicity, inhibits colony formation by myeloid precursor cells in vitro. Compared to cytosine-arabinoside this cytotoxicity is roughly one third.Supported by grants of the Swiss Cancer League (FOR 101.AK.77.2) and the Swiss Science Foundation (3.3320.74).Acknowledgments. We wish to thank Prof. A. Lévy for support in providing normal bone marrow samples and Miss Beatrice Rubin for skilled technical assistance.     

Granulocyte colony formation in vitro: Enhancement by human placental (umbilical cord) serum

Summary Addition of human placental umbilical cord serum to bone marrow cultures reproducibly increased the number of granulocyte colonies in vitro. This stimulatory effect was significantly greater than that of fetal calf serum which was seen in cultures of human bone marrow under the conditions described.This work was financed by grant FOR. No. 004.AK.71 (1) from the Swiss Cancer League.The authors wish to express their gratitute to colleagues from Maternity Ward, Frauenspital, Bern, and Department of Thoracic and Vascular Surgery, Inselspital, Bern, through whose kind assistance samples of umbilical blood and fragments of ribs removed at thotacotomies were obtained. MissJ. Blom provided excellent technical assistance.     

小型猪骨髓间充质干细胞经心导管冠状动脉移植及其迁移和分化的研究

目的建立小型猪心导管介入冠状动脉治疗模型,为骨髓间充质于细胞（MSCs）移植治疗心血管疾病提供新的方法,观察骨髓间充质干细胞经心导管介入冠状动脉移植后在心肌内的迁移及分化。方法选用冠状动脉解剖生理特点与人类相似的小型猪,应用心导管介入技术将体外培养扩增的小型猪自体MSCs移植进入左冠状动脉前降支,用免疫荧光检测即刻移植和移植后6W移植细胞的肌钙蛋白T（cTnT）、缝隙连接蛋白43（Cx43）、anti－Ⅷfactor的表达。结果成功的完成了小型猪心导管介入冠状动脉进行自体骨髓间充质干细胞移植。移植细胞存活,向心肌组织迁移,并向心肌细胞和毛细血管方向分化。结论该方法稳定,技术先进。可进行准确的定位和动态观测,为临床应用提供了一个可靠的技术平台。MSCs移植后在心肌内发生迁移及分化。     

Factor(s) from nonmacrophage bone marrow stromal cells inhibit Lewis lung carcinoma and B16 melanoma growth in mice

Bone marrow stroma produces positive and negative growth regulators which constitute the hematopoietic microenvironment. As many tumors metastasize to the bones, these regulators may also influence tumor growth. Hematopoietic cytokines may indeed exert both positive and negative effect on tumor growth. We report that, when mixed with tumor cells. adherent bone marrow cells inhibit primary tumor growth and metastases formation in mice transplanted with Lewis lung carcinoma or B16 melanoma. Peritoneal macrophages or lymph node cells did not exert any influence. The tumor inhibition was apparently due to soluble factor(s) released by marrow stromal cells. In cocultures with B16 melanoma cells, adherent bone marrow cells exerted a significant antiproliferative effect which was increased by previous culture of the bone marrow cells with granulocyte-macrophage colony-stimulating factor but not with macrophage colony-stimulating factor. Neither neutralizing antibodies against tumor necrosis factor-alpha, transforming growth factor-beta or interferon alpha/beta nor addition of Escherichia coli lipopolysaccharide to generate inflammatory cytokines could affect the antiproliferative effect of bone marrow stromal cells. The bone marrow stroma factor(s) which inhibit tumor growth might, therefore, be a novel growth regulator.     

Age-related differences in the effect of in vivo administration of indomethacin on hemopoietic cell lineages of the spleen and bone marrow of mice

During 21 days of indomethacin treatment, erythroid cells in the spleens of both young adult and older mice, and in the bone marrow of young adult mice, were increased significantly early, in treatment, relative to age-matched control organs, and remained high throughout treatment. During drug exposure, the numbers of myeloid cells in young adult bone marrow, but not spleen, were reduced, but in older mice these cells were elevated in both organs. Lymphoid cells in the young adult and older mouse spleens decreased and increased, respectively, during treatment, but were unchanged and decreased, respectively, in the bone marrow of young adult and older mice. Monocytemacrophage cells in the spleen were elevated but unchanged in the bone marrow of both age groups. During 14 days of indomethacin treatment of houng adult mice, the proportions of precursor cells in DNA synthesis of only the splenic erythroid lineage were increased. Thus, the major hemopoietic lineages in both the bone marrow and spleen are affected by exposure to indomethacin in a time-dependent and age-dependent manner. For all lineages studied, those of the bone marrow were least disturbed, and/or were first to recover, even during continued drug exposure.     

Age-related differences in the effect of in vivo administration of indomethacin on hemopoietic cell lineages of the spleen and bone marrow of mice.

During 21 days of indomethacin treatment, erythroid cells in the spleens of both young adult and older mice, and in the bone marrow of young adult mice, were increased significantly early in treatment, relative to age-matched control organs, and remained high throughout treatment. During drug exposure, the numbers of myeloid cells in young adult bone marrow, but not spleen, were reduced, but in older mice these cells were elevated in both organs. Lymphoid cells in the young adult and older mouse spleens decreased and increased, respectively, during treatment, but were unchanged and decreased, respectively, in the bone marrow of young adult and older mice. Monocyte-macrophage cells in the spleen were elevated but unchanged in the bone marrow of both age groups. During 14 days of indomethacin treatment of young adult mice, the proportions of precursor cells in DNA synthesis of only the splenic erythroid lineage were increased. Thus, the major hemopoietic lineages in both the bone marrow and spleen are affected by exposure to indomethacin in a time-dependent and age-dependent manner. For all lineages studied, those of the bone marrow were least disturbed and/or were first to recover, even during continued drug exposure.     

Alterations in bone-marrow cellularity following thymectomy in rats.

The number of nucleated marrow cells was decreased following neonatal thymectomy in rats, and was corrected by administration of syngeneic lymphoid cells, or by implantation of a syngeneic testis. These results suggest that, in the rat, as has been shown previously in the mouse, lymphoid cells exert parital control over bone marrow cellularity and this effect may be further modulated by sex steroids.     

Secretion of a granular colony-stimulating factor by an adherent layer of long-term hematopoietic bone marrow cultures in mice

The presence of a CSF activity in long term bone marrow cultures in Mice was not described until now. Using a double layer agar technique directly on the adherent layers of the cultures, a strong CSF activity is detected in these adherent layers, before recharging the cultures at the third week, or when the cultures are not recharged. The role of this activity in long term myelopoiesis maintenance in vitro is discussed.     

Regulation of neutrophil trafficking from the bone marrow

Neutrophils are an essential component of the innate immune response and a major contributor to inflammation. Consequently, neutrophil homeostasis in the blood is highly regulated. Neutrophil number in the blood is determined by the balance between neutrophil production in the bone marrow and release from the bone marrow to blood with neutrophil clearance from the circulation. This review will focus on mechanisms regulating neutrophil release from the bone marrow. In particular, recent data demonstrating a central role for the chemokines CXCL12 and CXCL2 in regulating neutrophil egress from the bone marrow will be discussed.     

Stimulation of hematopoietic stem cells by antilymphocyte serum]

The physiopathogeny of aplastic anemia is still unknown, it can be related to a stem cell defect or a microenvironment disease. An autoimmune origin is suspected but not as yet proved. To demonstrate the autoimmune origin of some cases of aplastic anemia, we have studied the effect of antilymphocyte globulin (ALG) on the hematopoiesis of aplastic patients by serial hematological and bone marrow investigation including blood counts, bone marrow cellularity, scanning with indium and technetium and granulocytic colonies in agar, 8 out of 17 patients had a response to ALG as shown by a rise of granulocytes and reticulocytes counts, increase of bone marrow cellularity and number of granulocytic colonies. This study tends to show that ALG has a stimulating effect on hematopoiesis in some cases of severe aplastic anemia.     

Preservation of hematopoietic stem cells by slow freezing and elimination of the heat of fusion]

Previous experiments, in dogs with fresh bone marrow or bone marrow frozen with a modified freezing system have demonstrated a 100% recovery of frozen stem cells, stored for periods up to 5 months. Five patients, three with drug resistant acute leukemia and two with metastic carcinomas, have been treated with a high dose combination chemotherapy regimen (TACC) followed by reinfusion of marrow cryopreserved with the same modified freezing system. Following the reinfusion of marrow, autologous engraftment was demonstrated on bone marrow aspiration between days 5 and 10.     

Experimental allergic encephalomyelitis in T-lymphocyte deficient rats

Summary Thymectomized, lethally irradiated rats reconstituted with syngeneic bone marrow were injected with rat brain in complete Freund adjuvant mixture. Both, they and sham-thymectomized, irradiated and bone marrow protected rats displayed a higher incidence of leg paralysis than normal non-irradiated animals. Thymectomy lowered the incidence of the disease.Supported by the Research Fund of Croatia (Zagreb).     

Alterations in bone-marrow cellularity following thymectomy in rats

Summary The number of nucleated marrow cells was decreased following neonatal thymectomy in rats, and was corrected by administration of syngeneic lymphoid cells, or by implantation of a syngeneic testis. These results suggest that, in the rat, as has been shown previously in the mouse, lymphoid cells exert parital control over bone marrow cellularity and this effect may be further modulated by sex steroids.Supported in part by NSF-RIAS 77-06922, NIH AM21137 and the Morseman Foundation.     

The protective effect of thiola against the genotoxic action of benzo(a)pyrene

Summary The protective effect of Thiola against the genotoxicity, induced by benzo(a)pyrene, in vitro and in vivo, was investigated. By association of Thiola to benzo(a)pyrene a significant decrease of the numerical and structural chromosome aberrations and a reduction of the incidence of c-mitoses has been obtained in human diploid cells, i.e. human embryonic lung fibroblasts of the cell-line ICP-23, and C₅₆Bl/6 mouse bone marrow cells.