Abstract:
:Characterization of hematopoietic cells and measurement of their proliferative potential is critical in many research and clinical applications. Because in vivo assay of human cells is not possible and xenogeneic assays are not yet routine, in vitro assays such as the long-term culture-initiating cell (LTC-IC) assay have been widely adopted. This study investigated LTC-IC assay linearity and reproducibility and resulting implications with respect to quantitation of primitive cell expansion. Measurement of secondary colony-forming cells (2 degrees CFCs) from 5-week cultures of bone marrow (BM) mononuclear cells (MNCs) showed that 2 degrees CFC frequency varied with assay plating density in a nonlinear fashion. The measured 2 degrees CFC frequency increased from 4.6 to 63.8 (per 10(5) MNCs) as assay plating density was decreased from 5 x 10(5) to 2 x 10(4) MNCs per well (P < 10(-6), n = 37). In contrast, assay of CD34-enriched cells was linear within the range studied. Assays of cells obtained from expansion cultures initiated with either MNCs or CD34-enriched cells were also nonlinear. Consequently, calculated 2 degrees CFC expansion ratios were ambiguous and dependent on the assay plating densities used. Limiting dilution analysis (LDA) results were also nonlinear, with LTC-IC frequency increasing from 8. 2 to 22.4 per 10(5) MNCs (P < 10(-4), n = 100) as assay plating densities were decreased. Despite the nonlinearity, 2 degrees CFC and LTC-IC assay results were consistent and reproducible over time with different samples and techniques and gave a semiquantitative indication of relative primitive cell frequency. Although CD34-enriched cells gave linear assay output, purification of cells for every assay is impractical. Therefore, exposure of cells to 5-fluorouracil (5-FU) was explored for improving assay linearity. Incubation of MNCs in 250 microg/mL 5-FU for 1 to 2 hours depleted accessory cells and resulted in a cell population that gave linear 2 degrees CFC readout. The 5-FU-resistant LTC-ICs accounted for 49% of the total LTC-IC population, adding the potential benefit of restricting assay measurement to more primitive noncycling LTC-ICs. Consequently, similar linear assay results can be obtained with either the bulk 2 degrees CFC or LDA LTC-IC methods after 5-FU, but multiple plating densities are nevertheless still required in both methods due to the greater than 100-fold range in primitive cell frequency present in normal human donor BM.
journal_name
Bloodjournal_title
Bloodauthors
Koller MR,Manchel I,Smith AKsubject
Has Abstractpub_date
1998-06-01 00:00:00pages
4056-64issue
11eissn
0006-4971issn
1528-0020journal_volume
91pub_type
杂志文章相关文献
BLOOD文献大全abstract::The adhesion of leukocytes to platelets deposited at the site of vascular injury may represent an important mechanism by which leukocytes contribute to hemostasis and thrombosis. In this study, we examined whether, in comparison with their distribution in circulating blood, certain leukocyte types are enriched at site...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1997-02-15 00:00:00
abstract::Lymphoepithelioid lymphoma (Lennert's lymphoma) was first described as a special variant of Hodgkin's disease. This lesion is characterized by a high percentage of epithelioid and T cells and rarely contains the classical Hodgkin's/Reed-Sternberg cells. Cytogenetic abnormalities indicate that Lennert's lymphoma is of ...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1986-09-01 00:00:00
abstract::Adenovirus infections occur in 5% to 21% of patients following stem cell transplantation (SCT), with an associated mortality of up to 50%. However, a lack of prospective studies has hampered further developments in the understanding and management of this infection in the posttransplantation setting. We prospectively ...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2002-02-0377
更新日期:2002-09-01 00:00:00
abstract::Acute graft-versus-host disease (aGVHD) remains a major complication of allogeneic hematopoietic stem cell transplant (alloHSCT), underscoring the need to further elucidate its mechanisms and develop novel treatments. Based on recent observations that microRNA-155 (miR-155) is up-regulated during T-cell activation, we...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2011-10-387522
更新日期:2012-05-17 00:00:00
abstract::To investigate the role of peritoneal mesothelial cells in regulating hematopoiesis, as well as inflammation, healing, and tissue regeneration processes, long-term cultures of peritoneal mesothelial cells from human endocavitarian fluids were established. The purity of the cell population was assessed by morphologic a...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1992-12-01 00:00:00
abstract::A simple and reliable cytogenetic test for Fanconi's anemia (FA) that is based on the hypersensitivity of FA cells to mitomycin C (MC) is described. Equal volumes of whole blood from a patient in whom the diagnosis of FA is suspected and from a normal person of the opposite sex are co-cultured in phytohemagglutinin-co...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1987-06-01 00:00:00
abstract::Non-Hodgkin lymphomas (NHLs) are the most common cancer to affect pet dogs. In contrast to the many genes whose mutation contributes to lymphomagenesis in humans, relatively little is known about the acquired genetic alterations that lead to canine B-cell lymphomas (cBCLs). We performed a survey of 84 canine NHL tumor...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2014-10-602714
更新日期:2015-02-05 00:00:00
abstract::Defective function of dendritic cells (DC) in cancer has been recently described and may represent one of the mechanisms of tumor evasion from immune system control. We have previously shown in vitro that vascular endothelial growth factor (VEGF), produced by almost all tumors, is one of the tumor-derived factors resp...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1998-12-01 00:00:00
abstract::Perfluorocarbons have shown promise as clinical blood substitutes. Although early experience in Japan with one such product--Fluosol-DA--has been uncomplicated, we observed an adverse pulmonary reaction in the first American patient to receive it and know of similar reactions in two other Americans so treated. Postula...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1982-06-01 00:00:00
abstract::The retinoblastoma (Rb) tumor suppressor plays important roles in regulating hematopoiesis, particularly erythropoiesis. In an effort to understand whether Rb function can be mediated by E2F transcription factors in a BM-derived hematopoietic system in mice, we uncovered a functional synergy between Rb and E2F8 to pro...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2011-10-388231
更新日期:2012-05-10 00:00:00
abstract::In an experimental model we demonstrated that endothelial cells of all organs are targets of the alloimmune reaction. Here, in 68 digestive biopsies, we found endothelial lesions by immunohistochemistry and ultrastructure in patients with severe acute graft-versus-host disease (GVHD). In contrast, no such endothelial ...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2003-05-1548
更新日期:2004-06-15 00:00:00
abstract::Cellular interleukin 6 (IL-6) is an important growth factor for Kaposi sarcoma- associated herpesvirus (KSHV)-associated neoplasms, which include human immunodeficiency virus (HIV)-related and -unrelated cases of Kaposi sarcoma (KS), primary effusion lymphoma (PEL), and multicentric Castleman disease (MCD). Increased ...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood.v99.2.649
更新日期:2002-01-15 00:00:00
abstract::In an international collaborative study, a central histologic review identified 891 patients with essential thrombocythemia, strictly defined by World Health Organization criteria. After a median follow-up of 6.2 years, 109 (12%) patients experienced arterial (n = 79) or venous (n = 37) thrombosis. In multivariable an...
journal_title:Blood
pub_type: 杂志文章,多中心研究
doi:10.1182/blood-2011-02-339002
更新日期:2011-06-02 00:00:00
abstract::Fanconi anemia (FA) is an autosomal recessive syndrome featuring diverse symptoms including progressive bone marrow failure and early occurrence of acute myeloid leukemia. Nine genetic subtypes have been described for FA (A, B, C, D1, D2, E, F, G, and L), all of which have been connected to distinct disease genes, exc...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2003-08-2915
更新日期:2004-04-01 00:00:00
abstract::The hereditary stomatocytoses are a series of dominantly inherited hemolytic anemias in which the permeability of the erythrocyte membrane to monovalent cations is pathologically increased. The causative mutations for some forms of hereditary stomatocytosis have been found in the transporter protein genes, RHAG and SL...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2010-12-326645
更新日期:2011-11-10 00:00:00
abstract::Pacritinib (SB1518) is a Janus kinase 2 (JAK2), JAK2(V617F), and Fms-like tyrosine kinase 3 inhibitor that does not inhibit JAK1. It demonstrated a favorable safety profile with promising efficacy in phase 1 studies in patients with primary and secondary myelofibrosis (MF). This multicenter phase 2 study further chara...
journal_title:Blood
pub_type: 杂志文章,多中心研究
doi:10.1182/blood-2013-02-484832
更新日期:2015-04-23 00:00:00
abstract::Tn polyagglutinability syndrome is an acquired condition where erythrocytes express Tn neo-antigen and become susceptible to hemagglutination by the naturally occurring anti-Tn present in normal sera. Early studies had indicated that O-linked N-acetyl galactosamine was the sole serologic Tn determinant, but more recen...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1992-11-01 00:00:00
abstract::A deficiency of adenosine deaminase, an enzyme important in purine nucleoside catabolism, is associated with a severe combined immunodeficiency disease in children. Inhibition of this enzyme in vitro and in vivo results in an impairment in lymphoblast proliferation. We have investigated the pharmacologic inhibition of...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1981-03-01 00:00:00
abstract::The induction of tumor cell differentiation represents an attractive strategy for the treatment of a wide range of malignancies. Differentiation of HL-60 promyelocytic leukemia cells towards neutrophils or monocytes has been shown to induce apoptotic cell death, which is inhibited by bcl-2 over-expression. However, th...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1996-05-01 00:00:00
abstract::Red blood cells deliver O(2) from the lungs to every cell in the human body. Reduced tissue oxygenation triggers increased production of erythropoietin by hypoxia-inducible factor 1 (HIF-1), which is a transcriptional activator composed of an O(2)-regulated alpha subunit and a constitutively expressed beta subunit. Hy...
journal_title:Blood
pub_type: 杂志文章,评审
doi:10.1182/blood-2009-05-189985
更新日期:2009-09-03 00:00:00
abstract::Sphingosine 1-phosphate (S1P), a bioactive lipid, is produced and stored in platelets and is released from activated platelets during blood coagulation activation. Thrombin, which is also generated during blood coagulation, has been shown to induce tissue factor (TF), the initiator of blood coagulation, in endothelial...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2002-11-3607
更新日期:2003-09-01 00:00:00
abstract::Primary myelofibrosis (PMF) is the most serious myeloproliferative disorder, characterized by clonal myeloproliferation associated with cytokine-mediated bone marrow stromal reaction including fibrosis and osteosclerosis. Current drug therapy remains mainly palliative. Because the NF-kappaB pathway is implicated in th...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2006-10-054502
更新日期:2007-07-01 00:00:00
abstract::The stress-activated protein/c-Jun N-terminal kinases (SAPK/JNK) have been shown to be activated by pro-inflammatory cytokines, as well as physical and chemical stresses. We now show that a variety of hematopoietic growth factors, including Steel locus factor (SLF), granulocyte-macrophage colony-stimulating factor (GM...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1997-05-01 00:00:00
abstract::Chronic graft-versus-host disease (cGVHD) is an increasingly common cause of morbidity and mortality in allogeneic stem cell transplantation (alloSCT). Relative to acute GVHD (aGVHD), much less is understood about cGVHD. Using the B10.D2 --> BALB/c murine cGVHD model, which shares critical pathologic features with hum...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2004-01-0328
更新日期:2004-09-01 00:00:00
abstract::Autoimmune hemolytic anemia (AIHA) is a disease in which autoantibodies against red blood cells (RBCs) lead to their premature destruction. Most clinically significant autoantibodies are of the immunoglobulin G (IgG) type, which leads primarily to the uptake and destruction of RBCs by splenic and hepatic macrophages. ...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2001-11-0061
更新日期:2003-01-15 00:00:00
abstract::Platelet-type von Willebrand disease (vWD) is a congenital bleeding disorder characterized by heightened ristocetin-induced platelet aggregation caused by abnormally high affinity between the platelet membrane glycoprotein (GP) Ib/IX complex and von Willebrand factor (vWF). Two distinct point mutations, Gly233 to Val ...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1997-07-15 00:00:00
abstract::Targeted disruption of the Friend leukemia integration 1 (Fli-1) proto-oncogene results in severe dysmegakaryopoiesis and embryonic lethality. We used morula-stage aggregation as a strategy to further clarify the hematopoietic defects of the Fli-1 gene-targeted mice. Analyses of lineage expression of Fli-1(+/-) and Fl...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2003-12-4345
更新日期:2005-01-01 00:00:00
abstract::Combinations of proinflammatory and procoagulant reactions are the unifying principle for a variety of disorders affecting the cardiovascular system. The factor XII-driven contact system starts coagulation and inflammatory mechanisms via the intrinsic pathway of coagulation and the bradykinin-producing kallikrein-kini...
journal_title:Blood
pub_type: 杂志文章,评审
doi:10.1182/blood-2017-04-569111
更新日期:2018-04-26 00:00:00
abstract::Precursor B-cell acute lymphoblastic leukemias (B-ALLs) have been shown to be oligoclonal at the Ig heavy-chain (IgH) gene level in up to 40% of cases by Southern blot hybridization. In contrast, oligoclonality as deduced from diversity of T-cell receptor (TcR)-delta gene rearrangements of the immature types (ie, V de...
journal_title:Blood
pub_type: 杂志文章
doi:
更新日期:1995-05-15 00:00:00
abstract::The risk for thrombosis is increased in patients with hereditary hydrocytosis, an uncommon variant of hereditary stomatocytosis. Erythrocytes from 2 patients with hydrocytosis were studied to gain insight into the mechanism of thrombosis in this disorder. Erythrocytes demonstrated abnormal osmotic scan ektacytometry a...
journal_title:Blood
pub_type: 杂志文章
doi:10.1182/blood-2001-12-0329
更新日期:2003-06-01 00:00:00