Hemopoietic Development Research Articles

Lipid Accumulation and Production of Colony-Stimulating Activity by the 266AD Cell Line Derived From Mouse Bone Marrow

The availability of cloned lines of bone marrow stromal cells could facilitate the analysis of their role in hemopoietic cell development. The 266AD cell line was isolated from a colony of lipid-accumulating bone marrow cells growing in a collagen gel. 266AD cells have subsequently been maintained by passage in tissue culture plastic flasks about every 10 days for greater than 10 mo. Subconfluent cultures of cells are fibroblast-appearing, but in confluent cell sheets, prominent foci of lipid-containing cells develop in both uncloned and four separate cloned cell lines. Supernatants from confluent cultures containing lipid-laden cells contain granulocyte-macrophage colony-stimulating activity (GM-CSA) for normal bone marrow cells and can induce differentiation of Abelson virus transformed murine promonocytic leukemia cells. 266AD cells were originally isolated in the presence of hydrocortisone, but hydrocortisone is not necessary for lipogenesis to occur. Growth of bone marrow cells in a collagen gel matrix provided a way to isolate stromal cells, and the 266AD cell line provides a means to examine the relationships between stromal cell lipogenesis and regulation of granulopoiesis.

Lipid accumulation and production of colony-stimulating activity by the 266AD cell line derived from mouse bone marrow

The availability of cloned lines of bone marrow stromal cells could facilitate the analysis of their role in hemopoietic cell development. The 266AD cell line was isolated from a colony of lipid-accumulating bone marrow cells growing in a collagen gel. 266AD cells have subsequently been maintained by passage in tissue culture plastic flasks about every 10 days for greater than 10 mo. Subconfluent cultures of cells are fibroblast-appearing, but in confluent cell sheets, prominent foci of lipid-containing cells develop in both uncloned and four separate cloned cell lines. Supernatants from confluent cultures containing lipid-laden cells contain granulocyte- macrophage colony-stimulating activity (GM-CSA) for normal bone marrow cells and can induce differentiation of Abelson virus transformed murine promonocytic leukemia cells. 266AD cells were originally isolated in the presence of hydrocortisone, but hydrocortisone is not necessary for lipogenesis to occur. Growth of bone marrow cells in a collagen gel matrix provided a way to isolate stromal cells, and the 266AD cell line provides a means to examine the relationships between stromal cell lipogenesis and regulation of granulopoiesis.

Book reviews

Clinical & Laboratory HaematologyVolume 2, Issue 3 p. 247-248 Book reviews First published: September 1980 https://doi.org/10.1111/j.1365-2257.1980.tb00830.xAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Abstract Iron Metabolism in Man, T.H. Bothwell, R.W. Charlton, J.D. Cooke & C.A. Finch Atlas of Human Hemopoietic Development E. Keleman, W. Calvo & T.M. Fliedner The Blood Picture and Urine Profile Edited by R.D.Neth & H.Aust. Volume2, Issue3September 1980Pages 247-248 RelatedInformation

Atlas of Human Hemopoietic Development.

Materials and Methods.- Diffuse Hemopoiesis.- Intravascular Hemopoietic Cells.- Organ-Bound Hemopoiesis.- References.- 5 Subject-Index.

Book Reviews

Book reviewed in this article:British Medical Bulletin, Chemical Carcinogenesis, vol. 36, no. 1 (Jan.), 1980. Scientific Editor P. BrookesDrug Carriers in Biology and Medicine. Edited by Gregory GregoriadisAtlas of Human Hemopoietic Development. By E. Kele‐men, W. Calvo and T. M. Fliedner

Murine hemopoietic colonies in culture containing normoblasts, macrophages, and megakaryocytes.

Murine marrow cells, when incubated in methylcellulose culture in the presence of erythropoietin and conditioned medium for two weeks, produced large macroscopic bursts containing normoblasts, macrophages, and often megakaryocytes. The clonal nature of these mixed colonies was supported by linearity studies and analysis of the percentages of constituent cells in different plating conditions. Time course observations and studies of the effects of L-cell-conditioned medium revealed that colony-forming cells for the mixed colonies (CFU-mix) are at earlier stages of hemopoietic development than burst-forming units (BFU-E). The mean of the modal sedimentation velocities of CFU-mix was 3.4 mm/hr and was in close agreement with that reported for the spleen colony-forming units. Almost none of the CFU-mix was in a DNA synthetic phase as measured by short-term exposure to tritiated thymidine. These results strongly indicate that CFU-mix represent a population of pluripotent hemopoietic stem cells in murine marrow.

Haemopoietic progenitor cells in prenatal congenitally anaemic 'flexed-tailed' (f/f) mice.

The incidences of erythroid colony forming cells (CFUe) and granulocyte-macrophage colony forming cells (CFUc) have been measured in 11-18 d prenatal livers of mice of genotype f/f and nearly congenic +/+ controls. In normal fetal livers numbers of CFUe (cells able to form colonies of 16 or more cells after 72 h in vitro) rise to a maximum on day 14 of gestation and represent c 1% of total fetal liver cells. In f/f fetal livers, peak values for numbers and proportions of CFUe are 50% of normal. The f/f lesion does not reduce the numbers of CFUc in fetal liver. Since this deficiency in CFUe parallels deficiencies of similar magnitude in spleen-colony forming units (CFUs) and erythroblasts in the liver, and erythrocytes in the blood, of f/f fetuses it is concluded that the f/f lesion is expressed at an early stage of haemopoietic development in prenatal life. The possibility that restricted haem synthesis is the primary effect of the f/f genotype and responsible for disturbances of both haemopoietic cellular proliferation and haemoglobin synthesis is examined.

Proliferation of hemopoietic stem cells and development of potentially leukemic cells in vitro.

Proliferation of hemopoietic stem cells and development of potentially leukemic cells in vitro.

Analysis of proliferation and differentiation of foetal granulocyte-macrophage progenitor cells in haemopoietic tissue.

ABSTRACTAnalysis of in vitro colony formation in agar cultures of foetal haemopoietic tissues of eight mammalian species has shown that granulocyte‐macrophage progenitor cells are present in foetal liver, yolk sac, marrow and spleen in numbers approaching the incidence in adult marrow. Such characteristics as buoyant density, growth rate and differentiation served to distinguish foetal from adult colony forming cells (CFCs).Cell cycle analysis performed by exposing haemopoietic cells to high doses of tritiated thymidine in vitro showed that foetal CFC proliferation in species of short gestation (rabbit, rat, mouse) approached or exceeded that observed in adult marrow. In contrast, in species of long gestation (human, monkey, calf, lamb, guinea‐pig) a period of variable duration was observed when foetal liver CFCs entered a non‐cycling G0 or blocked G1 phase. In these species foetal liver CFCs were found to be proliferating actively early in gestation and following the non‐cycling phase again re‐entered a proliferative state associated with onset of active granulopoiesis in foetal marrow and possible migration of CFC from liver to marrow.These results indicate the existence of granulocyte‐macrophage progenitor populations displaying foetal characteristics and adapted to particular stages of haemopoietic development, a situation which closely parallels that reported for erythropoiesis.