In vitro growth of rat bone marrow BFU-E.
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The erythropoietic response of Strain A mice with benzo(a)pyrene-induced fibrosarcoma has been studied. The rate of erythropoiesis, expressed in terms of 59Fe incorporation into circulating erythrocytes, was slightly increased in the fibrosarcomatous mice compared to the matched controls. The femoral marrow of the tumor hosts became hypocellular with reduced number of erythroblasts. The spleen, on the other hand, was hypercellular with an increased number of erythroid progenitor cells. Quantitative assessment of erythropoietic activity in the hematopoietic organs by measuring the 6-hr organ uptake of 59Fe revealed erythropoietic suppression in the marrow but enhancement in the spleen following tumor development. While the number of CFU-s in the femoral marrow of the tumor-bearing animals was slightly increased, marked increase in the concentration as well as absolute number of CFU-s was found in the spleen. Bioassay for erythropoietin revealed appreciable increase in the level of plasma erythropoietin in the tumor-bearing animals.
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Microenvironmentally dependent effects on murine haemopoiesis by a prolonged interleukin‐1 treatment
We administered recombinant human IL-1 beta (400 ng/d, s.c.) for 10 d to normal C57B1 mice and determined daily granuloid and erythroid parameters in marrow, spleen and blood. In the marrow CFU-GM numbers were not affected but later granuloid cell stages were moderately enhanced (170%). In the spleen, however, CFU-GM numbers were sharply increased (1600%), whereas the granuloid precursors only doubled. Blood granulocytes increased transiently to 275% on day 5. In the marrow all erythroid parameters were severely reduced. This reduction was partially compensated by the spleen where initially only BFU-E and with some delay also more mature erythroid cells accumulated. At the end of the treatment mice were slightly anaemic. When mice were treated with IL-1 and erythropoietin (10 U/d) simultaneously, the inhibitory effects on erythropoiesis were less severe. In agreement with in vivo results, IL-1 inhibited in vitro colony growth of CFU-E from normal bone marrow and spleen but spleen CFU-E from 5 d IL-1 treated mice were insensitive. We conclude that IL-1 can induce stimulation or inhibition of haemopoietic progenitor cells depending on their microenvironment.
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Infection of mice with the polycythemia-inducing strain of Friend leukemia virus caused a rapid emergence of new erythroid precursor cells. These cells which, in the absence of erythropoietin, proliferated in vitro to form colonies and even differnetiated, quickly out-numbered the usual erythropoietin-dependent hematopoietic elements in bone marrow and spleen. Ultimately, the marrow and spleen were probably totally repopulated with this erythropoietin-independent cell.
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S ummary . Three assays for erythropoietic progenitor cells have been applied to mice of genotype f/f and to nearly congenic +/+ controls. When f/f mice were tested for their ability to generate transient endogenous erythroid spleen colonies 4–6 days after 800 rads and 10 units of erythropoietin, the numbers of such colonies detected were greatly reduced, although normal numbers of spleen colonies appeared at later times (9–12 days) postirradiation. In contrast, cells capable of erythropoietic colony formation in culture (CFU‐E) were present within the normal range in both f/f spleen and marrow and their sensitivity to erythropoietin in culture was the same as that found previously for CFU‐E in the marrow and spleen of +/+ mice. Transfusion‐induced plethora reduced the number of CFU‐E in marrow to a similar extent in both f/f and +/+ mice; likewise, subsequent administration of 10 units of erythropoietin induced a rapid return in the number of marrow CFU‐E in both genotypes. In the spleen, CFU‐E numbers were approximately three‐fold lower in f/f mice in each group. These results support the view that the 5 day assay for transient endogenous spleen colonies detects cells (TE‐CFU) that are different from both CFU‐E and pluripotent stem cells (CFU‐S), although possibly overlapping to some extent with the immediate progenitors of CFU‐E. The results also indicate that the generation or maturation of TE‐CFU represents a primary site of expression of the f/f defect.
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