Biochemical and phenotypic characterization of human basophilic cells derived from dispersed fetal liver with murine T cell factors.
Metachromatically granulated cells were generated from human fetal liver stem cells cultured in heterologous mouse conditioned medium rich in interleukin 3. After 2 to 3 wk of culture with biweekly changes of medium and selection of nonadherent cells, all cells present in five cultures had cytoplasmic granules, and 60 to 95% of the cells stained metachromatically with toluidine blue or with alcian blue but not with the safranin counterstain. Ultrastructurally, many granules contained fibrillar material or electron-dense cores with fibrils and vesicular fragments. In addition, the granules of many cells were filled with electron-dense material, which in some cases had a fine structure consisting of concentric whorls or a reticular pattern. Analysis of high-affinity IgE receptors on the cultured cells by flow cytometry demonstrated a unimodal fluorescence pattern, suggesting that most cells were in the basophil or mast cell lineage. The cultured cells lacked the lymphoid cell surface determinants B1, B4, T3, and T11, the myeloid determinants Mo2 and MY9, the natural killer cell determinant 901, and Ia histocompatibility antigens, but expressed the myeloid determinant MY7. The cells contained 52 ng/10(6) cells of histamine and incorporated [35S]sulfate at an average rate of 31,300 cpm/10(6) cells/4 hr into 175,000 m.w. chondroitin sulfate A proteoglycans. Upon activation with 1 microM calcium ionophore A23187, the cultured cells released 53% of their cell-associated histamine and metabolized arachidonic acid to 15.0 ng/10(6) cells of immunoreactive leukotriene C4 equivalents, 0.5 ng/10(6) cells of leukotriene B4, and 3.1 ng/10(6) cells of prostaglandin D2 (means, n = 3). Thus, stem cells present in human fetal liver give rise, as do stem cells in mouse fetal liver, to metachromatically granulated cells when cultured in the presence of mouse interleukin 3. In both species, the cultured cells bear IgE receptors, lack characteristic lymphoid and most myeloid cell surface determinants, and contain histamine and chondroitin sulfate proteoglycans. The human fetal liver-derived cells are similar in morphology and T cell factor dependence to basophil-like cells derived from umbilical cord blood, but are novel in their capacity to generate leukotrienes and prostaglandin D2.
Seldin
D C
DC
Caulfield
J P
JP
Hein
A
A
Osathanondh
R
R
Nabel
G
G
Schlossman
S F
SF
Stevens
R L
RL
Austen
K F
KF
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D001491:Basophils / Q000166:cytology / Q000502:physiology*; D000001:Calcimycin / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D005434:Flow Cytometry; D006636:Histamine Release; D006801:Humans; D007377:Interleukin-3; D008099:Liver / Q000166:cytology / Q000196:embryology; D008222:Lymphokines / Q000494:pharmacology; D051379:Mice; D008854:Microscopy, Electron; D011509:Proteoglycans / Q000032:analysis; D011961:Receptors, Fc / Q000378:metabolism; D017455:Receptors, IgE; D011971:Receptors, Immunologic / Q000378:metabolism; D013601:T-Lymphocytes / Q000502:physiology
D000954:Antigens, Surface; D001095:Arachidonic Acids; D007377:Interleukin-3; D008222:Lymphokines; D011509:Proteoglycans; D011961:Receptors, Fc; D017455:Receptors, IgE; D011971:Receptors, Immunologic; D016718:Arachidonic Acid; D000001:Calcimycin