Characteristics of in vitro colony formation by cells from haemopoietic tissues

Summary The characteristics of stimulation of colony formationin vitro from cells of mouse haemopoietic tissues has been briefly reviewed. Mouse kidney or embryo feeder cells, media conditioned by the cells from these tissues, normal or leukemic mouse sera, sera from leukemic or infectious mononucleosis patients, human urine and mouse embryo extracts are all sources of colony stimulating activity and their properties have been described. All sources of colony-stimulating activity produce clones of cells of the granulocyte series. In tritiated thymidine treated mice injection of preparations rich in colony-stimulating activity has been shown to produce a neutrophil leucocytosis and accelerate the rate of accumulation of labelled neutrophils in the blood. It is suggested that thein vitro assay can detect factors capable of stimulating granulocyte development.     

Stimulation of murine hemopoietic colony formation by human IL-6

A novel hemopoietic CSF has been identified in the medium conditioned by lectin-stimulated human T cells. The cDNA clone encoding this factor, isolated by functional expression cloning in monkey cos-1 cells, proved to be identical with the cDNA encoding the cytokine B cell stimulatory factor-2/IFN-beta 2, a factor now known as IL-6. In the murine system, IL-6 indirectly supports the formation of several different types of hemopoietic colonies, including those derived from early blast cells, and directly supports the proliferation of granulocyte/macrophage progenitors. These results expand the range of known target cells of IL-6 to include hemopoietic progenitors in addition to B cells, T cells, and fibroblasts and provide further evidence that this cytokine plays an important role within a network of interacting cytokines that regulates many different biologic responses.     

Stimulation of murine bone-marrow colony formation by conditioned medium from human fetal liver cells

Summary A substance that stimulates growth of colonies of mononuclear granulocytic cells derived from the bone marrow of mice was produced by incubating fetal liver cells (conditioned medium). This substance appears to have the same properties described elsehwere as colony-stimulating factor (CSF). The enhanced stimulatory ability of the conditioned medium fromhuman fetal liver cells compared to medium not conditioned suggests that fetal liver is a potent source of colony-stimulating factor.     

Stimulation of murine bone-marrow colony formation by conditioned medium from human fetal liver cells.

A substance that stimulates growth of colonies of mononuclear granulocytic cells derived from the bone marrow of mice was produced by incubating fetal liver cells (conditioned medium). This substance appears to have the same properties described elsewhere as colony-stimulating factor (CSF). The enhanced stimulatory ability of the conditioned medium from human fetal liver cells compared to medium not conditioned suggests that fetal liver is a potent source of colony-stimulating factor.     

Modification of exogenous haemopoietic spleen colony formation in irradiated mice by antisera against mouse thymocytes and mouse brain.

Treatment of donor bone marrow in vitro with both anti-thymocyte serum (ATS) and anti-mouse brain serum (RAMBS) inhibits the formation of haemopoietic spleen colonies in irradiated and reconstituted mice. This activity of the antisera may be completely (ATS) or partly (RAMBS) eliminated by absorption with thymocytes. The effect of the antisera is complement-independent. Most likely it depends on the existence and functionality of phagocytes (macrophages) in the recipients.     

Stimulation by normal and leukemic mouse sera of colony formation in vitro by mouse bone marrow cells

Using a modification of the agar gel method for bone marrow culture, serum from various strains of mice has been tested for colony stimulating activity. Ninety percent of sera from AKR mice with spontaneous or transplanted lymphoid leukemia and 40–50% of sera from normal or preleukemic AKR mice stimulated colony formation by C57B1 bone marrow cells. Sera from 6% of C3H and 30% of C57B1 mice stimulated similar colony formation. The incidence of sera with colony stimulating activity rose with increasing age. All colonies were initially mainly granulocytic in nature but later became pure populations of mononuclear cells. Bone marrow cells exhibited considerable variation in their responsiveness to stimulation by mouse serum. Increasing the serum dose increased the number and size of bone marrow cell colonies and with optimal serum doses, 1 in 1000 bone marrow cells formed a cell colony. Preincubation of cells with active serum did not stimulate colony formation by washed bone marrow cells. The active factor in serum was filterable, non-dialysable and heat and ether labile.     

Behaviorally conditioned suppression of the immune response by antilymphocyte serum

Previous studies have shown that cyclophosphamide, a drug with a broad spectrum of cytotoxic activity and one that produces noxious gastrointestinal side effects, can elicit taste aversion conditioning when paired with a non-immunosuppressive oral stimulus (saccharin) resulting in immunosuppression after subsequent exposure to the paired stimulus (1). The study reported here indicates that rabbit anti-rat lymphocyte serum (ALS) which is selectively cytotoxic for lymphocytes and does not produce sensory side effects can similarly induce taste aversion conditioning of the immune response. Rats were exposed to oral saccharin paired with ALS injection. Upon subsequent reexposure to saccharin alone the immunosuppressive effects of ALS were reenlisted and the primary mixed lymphocyte culture response of conditioned rats to allogeneic lymphocytes was suppressed by 35% compared to controls. The demonstrated influence of psychologic factors on the immune response has far reaching implications, especially to human medicine, and their role in the course of disease and recovery in man demands further investigation.     

Stimulation of heme accumulation and erythroid colony formation in cultures of chick bone marrow cells by chicken plasma

A fibrin clot culture system with high plating efficiency is described for the growth of erythroid cells from chick bone marrow. Erythroid colonies grown in the absence of adult chicken plasma (spontaneous colonies) were either benzidine-negative or weakly benzidine-positive. Colonies grown in the presence of chicken plasma were 90% strongly benzidine-positive and 40% more abundant than spontaneous colonies. Plasma from anemic chickens was more effective than control plasma in inducing heme accumulation (heme-stimulating activity) and in increasing the number of erythroid colonies (colony-stimulating activity). Spontaneous colonies from 48-h cultures were transformed into benzidine-positive colonies by exposing them for 6-10 h to chicken plasma.