Abstract：
:Unlike cell lines, human hematopoietic stem cells (HSCs) are less efficiently transduced with HIV-1 vectors, potentially limiting this approach. To investigate which step (internalization, reverse transcription, nuclear transport, and integration) limits lentiviral transduction, we evaluated the kinetics of lentiviral transduction in human CD34(+) cells. We transduced HeLa and CD34(+) cells with self-inactivating HIV-1 vector at low and tenfold higher multiplicity of infection (MOI) and evaluated vector amounts at various time points based on the rationale that if a given step was not limiting, tenfold greater vector amounts would be obtained at the tenfold higher MOI. We observed slower internalization (>60 min), a peak in reverse transcription at 24 hours, and completion of integration at 3 days in CD34(+) cells. In HeLa cells, there were approximately tenfold greater amounts at high MOI at all time points. When compared with HeLa cells, CD34(+) cells exhibited larger differences in vector amounts between high and low MOIs at 2-6 hours and a smaller difference at 12 hours to 10 days, revealing a limitation in human CD34(+) cell transduction around 12 hours, which corresponds to reverse transcription. In serial measurements of reverse transcription at 24 hours, vector amounts did not decrease once detected among CD34(+) cells. When using an HSC expansion medium, we observed less limitation for starting reverse transcription and more efficient transduction among CD34(+) cells in vitro and in xenografted mice. These data suggest that it is the initiation of reverse transcription that limits lentiviral transduction of human CD34(+) cells. Our findings provide an avenue for optimizing human CD34(+) cell transduction.
journal_name
Exp Hematoljournal_title
Experimental hematologyauthors
Uchida N,Green R,Ballantine J,Skala LP,Hsieh MM,Tisdale JFdoi
10.1016/j.exphem.2015.10.003subject
Has Abstractpub_date
2016-02-01 00:00:00pages
106-15issue
2eissn
0301-472Xissn
1873-2399pii
S0301-472X(15)00694-3journal_volume
44pub_type
杂志文章abstract：OBJECTIVE:Osteoblasts play an important role in regulating hematopoiesis in the bone marrow. Here we show that U2-OS, a widely used osteoblastic cell line derived from an osteosarcoma, has the capacity to support proliferation of human hematopoietic progenitor cells in vitro. In this study, U2-OS cells are characterize...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/s0301-472x(00)00127-2
更新日期：2000-04-01 00:00:00
abstract：:Transfusion with 10,000 or 20,000 marrow cells resulted in 30+ days survival of 15%-50% of mice exposed to an Ld90 or LD100 or radiation. The use of congenic mice with alloenzyme markers permitted the identification of host and donor cells in the peripheral blood of transfused animals. Donor cells were present initial...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1988-08-01 00:00:00
abstract：:An increase in triglyceride synthesis has been observed in cultures of human bone marrow fibroblasts after the cells reach confluence. The addition of hydrocortisone further enhances triglyceride synthesis. Conditioned medium from confluent cultures also stimulates adipogenesis, probably mediated by a factor released ...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1987-03-01 00:00:00
abstract：:Fanconi anemia (FA) is a rare recessive disease characterized by progressive bone marrow failure, congenital abnormalities, and increased incidence of cancers. To date, mutations in 22 genes can cause FA or an FA-like phenotype. In China, in addition to clinical information, FA diagnosis primarily relies on genetic se...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2018.07.003
更新日期：2018-10-01 00:00:00
abstract：:Fractionated whole body irradiation induces thymic lymphomas in C57 BL/Ka mice after 6-12 months. A graft of normal congenic bone marrow cells immediately after the last irradiation prevents the development of lymphomas by inducing the disappearance of preleukemic cells. When such a graft is performed one month later,...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1997-02-01 00:00:00
abstract：:Using an accurate technique for staining and scoring soft agar cultures, this study defines the incidence of circulating CFUEOS in normal donors. With whole mononuclear cells as the source of colony stimulating factor (CSF) and fetal calf serum, 49.6% +/- 3.5 of the colonies were eosinophilic; with human placental con...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1982-03-01 00:00:00
abstract：:Peripheral blood and bone marrow specimens from patients with polycythemia vera (PV) and chronic myelogenous leukemia (CML) were assayed for erythroid and granulopoietic progenitor cells. All compartments were increased in CML patients in relapse although the ratio of BFU-E to CFU-C numbers remained constant in all CM...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1980-01-01 00:00:00
abstract：OBJECTIVE:Podocalyxin expression on Ter119(+) erythroblasts is induced following administration of erythropoietin (Epo) or phenylhydrazine treatment, but is notably absent on committed erythroid progenitors during homeostatic red cell turnover. Following high-dose Epo administration in vivo, podocalyxin surface express...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2008.09.006
更新日期：2009-01-01 00:00:00
abstract：:With a new labeling technique in radiation chimeras, an attempt was made to determine the duration of the phases of the stem cell cycle including shortest and mean generation time and to estimate the number of hemopoietic stem cells per unit of bone marrow cellularity. The DNA of bone marrow cells in DNA synthesis was...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1979-10-01 00:00:00
abstract：:The effects of a single injection of hemin on murine marrow BFU-E and CFU-S were assessed to determine whether hemin is as effective in augmenting primitive (day 7) BFU-E levels in situ as it is in vitro and to assess hemin's action on transplantable pluripotent stem cells (CFU-S). The results show that hemin exerts a...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1984-11-01 00:00:00
abstract：:The effect of recombinant interleukin 3 (IL-3) on the function of murine resident peritoneal macrophages was investigated. IL-3 enhanced the phagocytosis of Candida pseudotropicalis and Candida albicans and enhanced killing of the former. The enhanced killing is inhibited by scavengers of oxygen radicals, suggesting t...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1989-09-01 00:00:00
abstract：:Ouabain has been shown to increase the number of clonally derived erythroid stem cells, CFUE and BFUE, from normal murine bone marrow. We report here that digoxin and theophylline also enhance erythroid stem cell colony formation, in the presence of suboptimal concentrations of erythropoietin (Ep) (0.01 IU/ml) in meth...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1982-09-01 00:00:00
abstract：:More than 2million human erythroblasts extrude their nuclei every second in bone marrow under hypoxic conditions (<7% O2). Enucleation requires specific signal transduction pathways and the local assembly of contractile actomyosin rings. However, the energy source driving these events has not yet been identified. We e...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2019.02.004
更新日期：2019-04-01 00:00:00
abstract：OBJECTIVE:Mice harboring c-Myb hypomorphic mutations display enhanced thrombopoiesis because of increased numbers of megakaryocytes and their progenitors. Thrombopoietin induces these same effects, which lead us to hypothesize that the hormone acts through modulation of c-Myb expression, as c-Myb levels falls during th...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2008.07.001
更新日期：2008-12-01 00:00:00
abstract：OBJECTIVE:The function of neutrophils as primary mediators of innate immunity depends on the activity of granule proteins and critical components of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. Expression of their cognate genes is regulated during neutrophil differentiation by a complex netw...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2008.02.009
更新日期：2008-07-01 00:00:00
abstract：:A single injection of > or =10 microg/kg PEG-rHuMGDF in mice causes a dose-dependent increase in circulating platelets beginning on day 3 and peaking on days 5-6. The mean platelet volume and platelet distribution width at doses > or =100 microg/kg initially increase in a dose-dependent fashion and later decrease. How...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/s0301-472x(98)00012-5
更新日期：1999-01-01 00:00:00
abstract：:Similarly to other tissues, the bone marrow contains subsets of resident tissue macrophages, which are essential to maintain bone formation, functional hematopoietic stem cell (HSC) niches, and erythropoiesis. Pharmacologic doses of granulocyte colony-stimulating factor (G-CSF) mobilize HSC in part by interfering with...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2014.03.009
更新日期：2014-07-01 00:00:00
abstract：:Granulocyte colony-stimulating factor (G-CSF) is used routinely in the clinical setting to mobilize hematopoietic stem progenitor cells (HSPCs) into the patient's blood for collection and subsequent transplantation. However, a significant proportion of patients who have previously received chemotherapy or radiotherapy...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2017.05.004
更新日期：2017-08-01 00:00:00
abstract：OBJECTIVE:Adult T-cell leukemia (ATL) is a mature CD4(+) T-cell malignancy caused by infection with human T-lymphotrophic virus type-1 and is associated with a marked hypercalcemia in many patients. Recently, it has been proposed that macrophage inflammatory protein-1alpha (MIP-1alpha) is the clinical hallmark of hyper...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2007.11.013
更新日期：2008-04-01 00:00:00
abstract：:Effects of thyroid hormones on the production of erythropoietin (Epo) were investigated in isolated perfused rat kidneys and in the human hepatoma cell line, HepG2. Epo protein was measured by radioimmunoassay. L-triiodothyronine and L-thyroxine stimulated hypoxia-induced Epo formation both in the kidney and in HepG2 ...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1994-03-01 00:00:00
abstract：:In slightly more than a decade of stem cell plasticity research, 24 peer-reviewed articles have demonstrated plasticity across organ and/or embryonic lineage boundaries at the single-cell level, with only 1 article showing negative results. These data, taken together with data about reversibility of gene restrictions ...
journal_title：Experimental hematology
pub_type： 杂志文章,评审
doi：10.1016/j.exphem.2010.04.013
更新日期：2010-07-01 00:00:00
abstract：:The effects of intraperitoneal Ehrlich ascites tumor (ET) growth on the kinetics of hemopoietic stem cells in the host bone marrow were studied using the spleen colony and the soft agar culture techniques. There is a decrease in spleen colony forming capacity of bone marrow of ET bearing mice, whereas in vitro assays ...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1978-05-01 00:00:00
abstract：:Two main aspects of the in vitro mouse foetal liver cell assay for Erythroid Stimulating Factor (ESF) in human sera have been investigated. The haem extraction process has been shown to give specific and quantitative recovery of 59Fe labelled haem from haemoglobin thus confirming that the material assayed in human ser...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1975-11-01 00:00:00
abstract：OBJECTIVE:Approximately 60% of all anaplastic large-cell lymphomas (ALCL) contain a specific t(2;5)(p23;q35) chromosomal translocation leading to overexpression of NPM-ALK. As the chimeric tyrosine kinase is involved in tumorigenesis and pathogenesis of ALCL, we were interested to inhibit NPM-ALK expression using an ex...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/s0301-472x(02)01084-6
更新日期：2003-03-01 00:00:00
abstract：:Colony formation by megakaryocyte (MK) progenitors was studied in 36 normal individuals and in 26 patients with primary thrombocythemia (PT) using an improved plasma clot cloning system. MK colonies were identified by immunoperoxidase staining using a monoclonal antibody against human platelet glycoprotein IIb/IIIa co...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1989-01-01 00:00:00
abstract：:A method is described that permits the removal of hematopoietic cells and macrophages from mouse bone marrow cultures. The method is based on the difference in effect of extracellular ATP4- ions (ATP in the absence of divalent, complexing cations) on cells of hematopoietic origin, including macrophages, and of nonhema...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1994-02-01 00:00:00
abstract：OBJECTIVE:Mechanisms of T-cell stimulation by Flt3 ligand (Flt3L) and granulocyte-macrophage colony-stimulating factor (GM-CSF) remain unclear. Herein, we compared the effects of Flt3L and GM-CSF on the expansion of dendritic cells (DC) and T-cell subsets and cytokine expression. METHODS:Naïve and effector/memory T ce...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/s0301-472x(01)00722-6
更新日期：2001-10-01 00:00:00
abstract：OBJECTIVE:Regulation of the cell cycle by cyclin-dependent kinase (CDK) activity occurs at multiple levels and is often altered in human cancers. Therefore, CDK activity has been targeted for drug discovery, and a number of small molecules have now been identified as CDK inhibitors. Plant cytokinin analogues with CDK i...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/s0301-472x(02)00894-9
更新日期：2002-10-01 00:00:00
abstract：:Erythropoiesis requires large amounts of iron for hemoglobin synthesis. There are two sources of iron for erythropoiesis, dietary and stored iron; however, their relative contributions to erythropoiesis remain unknown. In this study, we used the stable iron isotope (57)Fe to quantify synthesis of hemoglobin derived fr...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：10.1016/j.exphem.2016.02.005
更新日期：2016-06-01 00:00:00
abstract：:Serum levels of monokines, including macrophage colony-stimulating factor (M-CSF), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha) and IL-1 beta (IL-1 beta), were measured in patients with chronic renal failure in an attempt to clarify the kinetics of these cytokines in the course of renal an...
journal_title：Experimental hematology
pub_type： 杂志文章
doi：
更新日期：1993-02-01 00:00:00
