High levels of lymphoid expression of enhanced green fluorescent protein in nonhuman primates transplanted with cytokine-mobilized peripheral blood CD34(+) cells.
We have used a murine retrovirus vector containing an enhanced green fluorescent protein complimentary DNA (EGFP cDNA) to dynamically follow vector-expressing cells in the peripheral blood (PB) of transplanted rhesus macaques. Cytokine mobilized CD34(+) cells were transduced with an amphotropic vector that expressed EGFP and a dihydrofolate reductase cDNA under control of the murine stem cell virus promoter. The transduction protocol used the CH-296 recombinant human fibronectin fragment and relatively high concentrations of the flt-3 ligand and stem cell factor. Following transplantation of the transduced cells,up to 55% EGFP-expressing granulocytes were obtained in the peripheral circulation during the early posttransplant period. This level of myeloid marking,however,decreased to 0.1% or lower within 2 weeks. In contrast,EGFP expression in PB lymphocytes rose from 2%-5% shortly following transplantation to 10% or greater by week 5. After 10 weeks,the level of expression in PB lymphocytes continued to remain at 3%-5% as measured by both flow cytometry and Southern blot analysis,and EGFP expression was observed in CD4(+),CD8(+),CD20(+),and CD16/56(+) lymphocyte subsets. EGFP expression was only transiently detected in red blood cells and platelets soon after transplantation. Such sustained levels of lymphocyte marking may be therapeutic in a number of human gene therapy applications that require targeting of the lymphoid compartment. The transient appearance of EGFP(+) myeloid cells suggests that transduction of a lineage-restricted myeloid progenitor capable of short-term engraftment was obtained with this protocol. (Blood. 2000;95:445-452)
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产品号#：
04436
04064
04100
04230
04236
04431
04434
04444
04464
04531
04535
04545
04536
04564
04035
04330
04034
04044
04435
04445
04534
04544
产品名：
MethoCult™ SF H4436
MethoCult™ H4034 Optimum 入门试剂盒
MethoCult™ H4100
MethoCult™ H4230
MethoCult™ SF H4236
MethoCult™ H4431
MethoCult™ H4434 Classic
MethoCult™ H4434 Classic
MethoCult™ H4434 Classic 套装
MethoCult™ H4531
MethoCult™ H4535 Enriched，不含EPO
MethoCult™ H4535 Enriched，不含EPO
MethoCult™ SF H4536
MethoCult™ H4534 Classic 无 EPO 入门试剂盒
MethoCult™ 不含EPO的H4035 Optimum
MethoCult™ H4330
MethoCult™ H4034 Optimum
MethoCult™ H4034 Optimum
MethoCult™ H4435 Enriched
MethoCult™ H4435 Enriched
MethoCult™ H4534 Classic（不含 EPO）
MethoCult™ H4534 Classic（不含 EPO）
Seshasayee D et al. (JUL 2000)
The Journal of biological chemistry 275 30 22969--77
Intron 1 elements promote erythroid-specific GATA-1 gene expression.
The zinc finger protein GATA-1 functions in a concentration-dependent fashion to activate the transcription of erythroid and megakaryocytic genes. Less is understood,however,regarding factors that regulate the GATA-1 gene. Presently elements within intron 1 are shown to markedly affect its erythroid-restricted transcription. Within a full-length 6. 8-kilobase GATA-1 gene construct (G6.8-Luc) the deletion of a central subdomain of intron 1 inhibited transcription textgreater/=10-fold in transiently transfected erythroid SKT6 cells,and likewise inhibited high-level transcription in erythroid FDCW2ER-GATA1 cells. In parental myeloid FDCER cells,however,low-level transcription was largely unaffected by intron 1 deletions. Within intron 1,repeated GATA and Ap1 consensus elements in a central region are described which when linked directly to reporter cassettes promote transcription in erythroid SKT6 and FDCER-GATA1 cells at high rates. Moreover,GATA-1 activated transcription from this subdomain in 293 cells,and in SKT6 cells this subdomain footprinted in vivo. For stably integrated GFP reporter constructs in erythroid SKT6 cells,corroborating results were obtained. Deletion of intronic GATA and Ap1 motifs abrogated the activity of G6.8-pEGFP; activity was decreased by 43 and 56%,respectively,by the deletion of either motif; and the above 1800-base pair region of intron 1 per se was transcribed at rates uniformly greater than G6.8-pEGFP. Also described is the differential utilization of exons 1a and 1b among primary erythromegakaryocytic and myeloid cells.
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产品号#：
产品名：
Wray J et al. (AUG 2009)
Blood 114 9 1852--8
Metnase mediates chromosome decatenation in acute leukemia cells.
After DNA replication,sister chromatids must be untangled,or decatenated,before mitosis so that chromatids do not tear during anaphase. Topoisomerase IIalpha (Topo IIalpha) is the major decatenating enzyme. Topo IIalpha inhibitors prevent decatenation,causing cells to arrest during mitosis. Here we report that acute myeloid leukemia cells fail to arrest at the mitotic decatenation checkpoint,and their progression through this checkpoint is regulated by the DNA repair component Metnase (also termed SETMAR). Metnase contains a SET histone methylase and transposase nuclease domain,and is a component of the nonhomologous end-joining DNA double-strand break repair pathway. Metnase interacts with Topo IIalpha and enhances its decatenation activity. Here we show that multiple types of acute leukemia cells have an attenuated mitotic arrest when decatenation is inhibited and that in an acute myeloid leukemia (AML) cell line this is mediated by Metnase. Of further importance,Metnase permits continued proliferation of these AML cells even in the presence of the clinical Topo IIalpha inhibitor VP-16. In vitro,purified Metnase prevents VP-16 inhibition of Topo IIalpha decatenation of tangled DNA. Thus,Metnase expression levels may predict AML resistance to Topo IIalpha inhibitors,and Metnase is a potential therapeutic target for small molecule interference.
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产品号#：
02690
09850
产品名：
StemSpan™ CC100
Tomihara K et al. (JUN 2010)
Journal of immunology (Baltimore,Md. : 1950) 184 11 6151--60
Antigen-specific immunity and cross-priming by epithelial ovarian carcinoma-induced CD11b(+)Gr-1(+) cells.
Both innate and adaptive immune systems are considered important for cancer prevention,immunosurveillance,and control of cancer progression. It is known that,although both systems initially eliminate emerging tumor cells efficiently,tumors eventually escape immune attack by a variety of mechanisms,including differentiation and recruitment of immunosuppressive CD11b(+)Gr-1(+) myeloid suppressor cells into the tumor microenvironment. However,we show that CD11b(+)Gr-1(+) cells found in ascites of epithelial ovarian cancer-bearing mice at advanced stages of disease are immunostimulatory rather than being immunosuppressive. These cells consist of a homogenous population of cells that morphologically resemble neutrophils. Moreover,like dendritic cells,immunostimulatory CD11b(+)Gr-1(+) cells can strongly cross-prime,augmenting the proliferation of functional CTLs via signaling through the expression of costimulatory molecule CD80. Adoptive transfer of these immunostimulatory CD11b(+)Gr-1(+) cells from ascites of ovarian cancer-bearing mice results in the significant regression of s.c. tumors even without being pulsed with exogenous tumor Ag prior to adoptive transfer. We now show for the first time that adaptive immune responses against cancer can be augmented by these cancer-induced granulocyte-like immunostimulatory myeloid (CD11b(+)Gr-1(+)) cells,thereby mediating highly effective antitumor immunity in an adoptive transfer model of immunity.
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EasySep™小鼠TIL（CD45）正选试剂盒
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线上讲座Streamline Your Tissue Processing Using the Automated STEMprep™ Tissue Dissociator发布日期: 06/12/2025
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