The Hedgehog receptor Patched controls lymphoid lineage commitment.
A first step in hematopoiesis is the specification of the lymphoid and myeloid lineages from multipotent progenitor cells in the bone marrow. Using a conditional ablation strategy in adult mice,we show that this differentiation step requires Patched (Ptch),the cell surface-bound receptor for Hedgehog (Hh). In the absence of Ptch,the development of T- and B-lymphoid lineages is blocked at the level of the common lymphoid progenitor in the bone marrow. Consequently,the generation of peripheral T and B cells is abrogated. Cells of the myeloid lineage develop normally in Ptch mutant mice. Finally,adoptive transfer experiments identified the stromal cell compartment as a critical Ptch-dependent inducer of lymphoid versus myeloid lineage commitment. Our data show that Ptch acts as a master switch for proper diversification of hematopoietic stem cells in the adult organism.
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产品类型：
产品号#：
03434
03444
19756
19756RF
产品名：
MethoCult™ GF M3434
MethoCult™ GF M3434
J. C.-H. Hsieh et al. (dec 2019)
Scientific reports 9 1 19917
The Prognostic Value of Circulating Tumor Cells in Asian Neuroendocrine Tumors.
Circulating tumor cells (CTC) play important roles in various cancers; however,few studies have assessed their clinical utility in neuroendocrine tumors. This study aimed to prospectively evaluate the prognostic value of CTC counts in Asian patients with neuroendocrine tumors before and during anti-cancer therapy. Patients who were diagnosed with unresectable histological neuroendocrine tumors between September 2011 and September 2017 were enrolled. CTC testing was performed before and during anti-cancer therapy using a negative selection protocol. Chromogranin A levels were also assessed. Univariate and multivariate Cox's proportional hazard model with forward LR model was performed to investigate the impact of independent factors on overall survival and progression-free survival. Kaplan-Meier method with log-rank tests were used to determine the difference among different clinicopathological signatures and CTC cutoff. The baseline CTC detection rate was 94.3{\%} (33/35). CTC counts were associated with cancer stages (I-III vs. IV,P = 0.015),liver metastasis (P = 0.026),and neuroendocrine tumor grading (P = 0.03). The median progression-free survival and overall survivals were 12.3 and 30.4 months,respectively. In multivariate Cox regression model,neuroendocrine tumors grading and baseline CTC counts were both independent prognostic factors for progression-free survival (PFS,P = 0.005 and 0.015,respectively) and overall survival (OS,P = 0.018 and 0.023,respectively). In Kaplan-Meier analysis,lower baseline chromogranin A levels were associated with longer PFS (P = 0.024). Baseline CTC counts are associated with the clinicopathologic features of neuroendocrine tumors and are an independent prognostic factor for this malignancy.
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产品类型：
产品号#：
15122
15162
产品名：
RosetteSep™人CD45去除抗体混合物
RosetteSep™人CD45去除抗体混合物
R. Liang et al. ( 2020)
Cell stem cell 26 3 359--376.e7
Restraining Lysosomal Activity Preserves Hematopoietic Stem Cell Quiescence and Potency.
Quiescence is a fundamental property that maintains hematopoietic stem cell (HSC) potency throughout life. Quiescent HSCs are thought to rely on glycolysis for their energy,but the overall metabolic properties of HSCs remain elusive. Using combined approaches,including single-cell RNA sequencing (RNA-seq),we show that mitochondrial membrane potential (MMP) distinguishes quiescent from cycling-primed HSCs. We found that primed,but not quiescent,HSCs relied readily on glycolysis. Notably,in vivo inhibition of glycolysis enhanced the competitive repopulation ability of primed HSCs. We further show that HSC quiescence is maintained by an abundance of large lysosomes. Repression of lysosomal activation in HSCs led to further enlargement of lysosomes while suppressing glucose uptake. This also induced increased lysosomal sequestration of mitochondria and enhanced the competitive repopulation ability of primed HSCs by over 90-fold in vivo. These findings show that restraining lysosomal activity preserves HSC quiescence and potency and may be therapeutically relevant.
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Song W et al. (OCT 2016)
Journal of Biomedical Materials Research - Part A 104 3 678--687
Efficient generation of endothelial cells from human pluripotent stem cells and characterization of their functional properties
Although endothelial cells (ECs) have been derived from human pluripotent stem cells (hPSCs),large-scale generation of hPSC-ECs remains challenging and their functions are not well characterized. Here we report a simple and efficient three-stage method that allows generation of approximately 98 and 9500 ECs on day 16 and day 34,respectively,from each human embryonic stem cell (hESC) input. The functional properties of hESC-ECs derived in the presence and absence of a TGF$$-inhibitory molecule SB431542 were characterized and compared with those of human umbilical vein endothelial cells (HUVECs). Confluent monolayers formed by SB431542(+) hESC-ECs,SB431542(-) hESC-ECs,and HUVECs showed similar permeability to 10,000 Da dextran,but these cells exhibited striking differences in forming tube-like structures in 3D fibrin gels. The SB431542(+) hESC-ECs were most potent in forming tube-like structures regardless of whether VEGF and bFGF were present in the medium; less potent SB431542(-) hESC-ECs and HUVECs responded differently to VEGF and bFGF,which significantly enhanced the ability of HUVECs to form tube-like structures but had little impact on SB431542(-) hESC-ECs. This study offers an efficient approach to large-scale hPSC-EC production and suggests that the phenotypes and functions of hPSC-ECs derived under different conditions need to be thoroughly examined before their use in technology development. This article is protected by copyright. All rights reserved.
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产品类型：
产品号#：
05850
05857
05870
05875
07923
27215
27250
27216
27217
27260
27270
36254
18098
18098RF
85850
85857
85870
85875
27845
27945
27840
27865
27940
27965
产品名：
Dispase (1 U/mL)
37µm可逆滤筛，小 (15 mL)
37µm可逆滤筛，大 (50 mL)
70µm可逆滤筛，小 (15 mL)
100µm可逆滤筛，小 (15 mL)
70µm可逆滤筛，大 (50 mL)
100µm可逆滤筛，大 (50 mL)
DMEM/F-12 with 15 mM HEPES
mTeSR™1
mTeSR™1
Kourjian G et al. (MAY 2016)
Journal of Immunology 196 9 3595--607
HIV Protease Inhibitor-Induced Cathepsin Modulation Alters Antigen Processing and Cross-Presentation.
Immune recognition by T cells relies on the presentation of pathogen-derived peptides by infected cells,but the persistence of chronic infections calls for new approaches to modulate immune recognition. Ag cross-presentation,the process by which pathogen Ags are internalized,degraded,and presented by MHC class I,is crucial to prime CD8 T cell responses. The original degradation of Ags is performed by pH-dependent endolysosomal cathepsins. In this article,we show that HIV protease inhibitors (PIs) prescribed to HIV-infected persons variably modulate cathepsin activities in human APCs,dendritic cells and macrophages,and CD4 T cells,three cell subsets infected by HIV. Two HIV PIs acted in two complementary ways on cathepsin hydrolytic activities: directly on cathepsins and indirectly on their regulators by inhibiting Akt kinase activities,reducing NADPH oxidase 2 activation,and lowering phagolysosomal reactive oxygen species production and pH,which led to enhanced cathepsin activities. HIV PIs modified endolysosomal degradation and epitope production of proteins from HIV and other pathogens in a sequence-dependent manner. They altered cross-presentation of Ags by dendritic cells to epitope-specific T cells and T cell-mediated killing. HIV PI-induced modulation of Ag processing partly changed the MHC self-peptidome displayed by primary human cells. This first identification,to our knowledge,of prescription drugs modifying the regulation of cathepsin activities and the MHC-peptidome may provide an alternate therapeutic approach to modulate immune recognition in immune disease beyond HIV.
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产品类型：
产品号#：
17952
17952RF
100-0696
19654
19654RF
产品名：
EasySep™人CD4+ T细胞分选试剂盒
RoboSep™ 人CD4+ T细胞分选试剂盒
EasySep™人CD4+ T细胞分离试剂盒
EasySep™ Direct 人 PBMC 分选试剂盒
RoboSep™ Direct 人 PBMC 分选试剂盒
Kang HM et al. (JAN 2018)
Nature biotechnology 36 1 89--94
Multiplexed droplet single-cell RNA-sequencing using natural genetic variation.
Droplet single-cell RNA-sequencing (dscRNA-seq) has enabled rapid,massively parallel profiling of transcriptomes. However,assessing differential expression across multiple individuals has been hampered by inefficient sample processing and technical batch effects. Here we describe a computational tool,demuxlet,that harnesses natural genetic variation to determine the sample identity of each droplet containing a single cell (singlet) and detect droplets containing two cells (doublets). These capabilities enable multiplexed dscRNA-seq experiments in which cells from unrelated individuals are pooled and captured at higher throughput than in standard workflows. Using simulated data,we show that 50 single-nucleotide polymorphisms (SNPs) per cell are sufficient to assign 97% of singlets and identify 92% of doublets in pools of up to 64 individuals. Given genotyping data for each of eight pooled samples,demuxlet correctly recovers the sample identity of<99% of singlets and identifies doublets at rates consistent with previous estimates. We apply demuxlet to assess cell-type-specific changes in gene expression in 8 pooled lupus patient samples treated with interferon (IFN)-β and perform eQTL analysis on 23 pooled samples.
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产品类型：
产品号#：
17853
17853RF
17854
17854RF
17858
17858RF
17952
17952RF
100-0699
100-0694
100-0696
产品名：
EasySep™人CD8正选试剂盒 II
RoboSep™ 人CD8正选试剂盒 II
EasySep™人CD19正选试剂盒II
RoboSep™ 人CD19正选试剂盒II
EasySep™人CD14正选试剂盒II
RoboSep™ 人CD14正选试剂盒II
EasySep™人CD4+ T细胞分选试剂盒
RoboSep™ 人CD4+ T细胞分选试剂盒
EasySep™人CD8阳性选择试剂盒II
EasySep™人CD14正选试剂盒II
EasySep™人CD4+ T细胞分离试剂盒
Q. Haas et al. ( 2019)
Cancer immunology research 7 5 707--718
Siglec-9 Regulates an Effector Memory CD8+ T-cell Subset That Congregates in the Melanoma Tumor Microenvironment.
Emerging evidence suggests an immunosuppressive role of altered tumor glycosylation due to downregulation of innate immune responses via immunoregulatory Siglecs. In contrast,human T cells,a major anticancer effector cell,only rarely express Siglecs. However,here,we report that the majority of intratumoral,but not peripheral blood,cytotoxic CD8+ T cells expressed Siglec-9 in melanoma. We identified Siglec-9+ CD8+ T cells as a subset of effector memory cells with high functional capacity and signatures of clonal expansion. This cytotoxic T-cell subset was functionally inhibited in the presence of Siglec-9 ligands or by Siglec-9 engagement by specific antibodies. TCR signaling pathways and key effector functions (cytotoxicity,cytokine production) of CD8+ T cells were suppressed by Siglec-9 engagement,which was associated with the phosphorylation of the inhibitory protein tyrosine phosphatase SHP-1,but not SHP-2. Expression of cognate Siglec-9 ligands was observed on the majority of tumor cells in primary and metastatic melanoma specimens. Targeting the tumor-restricted,glycosylation-dependent Siglec-9 axis may unleash this intratumoral T-cell subset,while confining T-cell activation to the tumor microenvironment.
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产品类型：
产品号#：
17953
17953RF
100-0710
产品名：
EasySep™人CD8+ T细胞分选试剂盒
RoboSep™ 人CD8+ T细胞分选试剂盒
EasySep™人CD8+ T细胞分选试剂盒
M. Reyes et al. (jan 2019)
Science advances 5 1 eaau9223
Multiplexed enrichment and genomic profiling of peripheral blood cells reveal subset-specific immune signatures.
Specialized immune cell subsets are involved in autoimmune disease,cancer immunity,and infectious disease through a diverse range of functions mediated by overlapping pathways and signals. However,subset-specific responses may not be detectable in analyses of whole blood samples,and no efficient approach for profiling cell subsets at high throughput from small samples is available. We present a low-input microfluidic system for sorting immune cells into subsets and profiling their gene expression. We validate the system's technical performance against standard subset isolation and library construction protocols and demonstrate the importance of subset-specific profiling through in vitro stimulation experiments. We show the ability of this integrated platform to identify subset-specific disease signatures by profiling four immune cell subsets in blood from patients with systemic lupus erythematosus (SLE) and matched control subjects. The platform has the potential to make multiplexed subset-specific analysis routine in many research laboratories and clinical settings.
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产品类型：
产品号#：
17853
17853RF
17858
17858RF
17951
17951RF
100-0699
100-0694
100-0695
产品名：
EasySep™人CD8正选试剂盒 II
RoboSep™ 人CD8正选试剂盒 II
EasySep™人CD14正选试剂盒II
RoboSep™ 人CD14正选试剂盒II
EasySep™人T细胞分选试剂盒
RoboSep™ 人T细胞分选试剂盒
EasySep™人CD8阳性选择试剂盒II
EasySep™人CD14正选试剂盒II
EasySep™人T细胞分选试剂盒
R. Gao et al. (may 2020)
Science advances 6 20 eaaz8411
Competition between PAF1 and MLL1/COMPASS confers the opposing function of LEDGF/p75 in HIV latency and proviral reactivation.
Transcriptional status determines the HIV replicative state in infected patients. However,the transcriptional mechanisms for proviral replication control remain unclear. In this study,we show that,apart from its function in HIV integration,LEDGF/p75 differentially regulates HIV transcription in latency and proviral reactivation. During latency,LEDGF/p75 suppresses proviral transcription via promoter-proximal pausing of RNA polymerase II (Pol II) by recruiting PAF1 complex to the provirus. Following latency reversal,MLL1 complex competitively displaces PAF1 from the provirus through casein kinase II (CKII)-dependent association with LEDGF/p75. Depleting or pharmacologically inhibiting CKII prevents PAF1 dissociation and abrogates the recruitment of both MLL1 and Super Elongation Complex (SEC) to the provirus,thereby impairing transcriptional reactivation for latency reversal. These findings,therefore,provide a mechanistic understanding of how LEDGF/p75 coordinates its distinct regulatory functions at different stages of the post-integrated HIV life cycles. Targeting these mechanisms may have a therapeutic potential to eradicate HIV infection.
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EasySep™小鼠TIL（CD45）正选试剂盒
科学海报Rapid Isolation of Highly Purified, Functional and Expandable Human Regulatory T Cells
产品手册循环肿瘤细胞的分离
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