SepMate™ is a tube that facilitates the isolation of PBMCs or specific cell types by density gradient centrifugation. SepMate™ tubes contain an insert that provides a barrier between the density gradient medium and blood. SepMate™ eliminates the need for careful layering of blood (or bone marrow or cord blood) onto the density gradient medium, and allows for fast and easy harvesting of the isolated mononuclear cells with a simple pour. SepMate™ is also compatible with RosetteSep™ enrichment cocktails, allowing isolation of specific cell types in less than 30 minutes.
SepMate™-15 is designed for processing samples from 0.5 to 5 mL of sample.
Contains <div>
Polypropylene tube containing an insert
Subtype Centrifugation Tubes
Cell Type B Cells, Dendritic Cells, Monocytes, Mononuclear Cells, NK Cells, T Cells, T Cells, CD4+, T Cells, CD8+, T Cells, Other Subsets, T Cells, Regulatory
Species Human
Sample Source Bone Marrow, Cord Blood, Whole Blood
Selection Method Negative
Application Cell Isolation
Brand SepMate
Area of Interest Chimerism, HLA, Immunology
Data Figures
Figure 1. Recovery of mononuclear cells (MNCs) from peripheral blood using SepMate™-50 versus standard density gradient centrifiguation. Recovery of MNCs from fresh and 48-hour post blood draw enriched by density gradient centrifugation with SepMate™ (purple) or without (grey). There was no significant difference in the recovery of MNCS with and without SepMate™.
Figure 2. Human CD4+ T Cell Isolation using SepMate™-50 and RosetteSep™ Human CD4+ T Cell Enrichment Cocktail
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
A Comprehensive Structure-Function Study of Neurogenin3 Disease-Causing Alleles during Human Pancreas and Intestinal Organoid Development. X. Zhang et al. Developmental cell 2019 aug
Abstract
Neurogenin3 (NEUROG3) is required for endocrine lineage formation of the pancreas and intestine. Patients with NEUROG3 mutations are born with congenital malabsorptive diarrhea due to complete loss of enteroendocrine cells, whereas endocrine pancreas development varies in an allele-specific manner. These findings suggest a context-dependent requirement for NEUROG3 in pancreas versus intestine. We utilized human tissue differentiated from NEUROG3-/- pluripotent stem cells for functional analyses. Most disease-associated alleles had hypomorphic or null phenotype in both tissues, whereas the S171fsX68 mutation had reduced activity in the pancreas but largely null in the intestine. Biochemical studies revealed NEUROG3 variants have distinct molecular defects with altered protein stability, DNA binding, and gene transcription. Moreover, NEUROG3 was highly unstable in the intestinal epithelium, explaining the enhanced sensitivity of intestinal defects relative to the pancreas. These studies emphasize that studies of human mutations in the endogenous tissue context may be required to assess structure-function relationships.
Oral administration of hydroxylated-graphene quantum dots induces intestinal injury accompanying the loss of intestinal stem cells and proliferative progenitor cells. L. Yu et al. Nanotoxicology 2019
Abstract
Graphene quantum dots (GQDs) have gained significant attention in various biomedical applications. The physicochemical properties of these nanoparticles, including toxic effects, are largely determined by their surface modifications. Previous studies have demonstrated high in vitro cytotoxicity of the hydroxylated GQDs (OH-GQDs). The focus of this study was on the intestinal toxicity of OH-GQDs. Briefly, C57BL/6J mice were given daily oral gavage of 0.05, 0.5 or 5 mg/kg OH-GQD for 7 days, and the indices of intestinal damage were evaluated. Higher doses of the OH-GQDs caused significant intestinal injuries, such as enhanced intestinal permeability, shortened villi and crypt loss. The number of Lgr5+ intestinal stem cells also decreased dramatically upon OH-GQDs exposure, which also inhibited the Ki67+ proliferative progenitor cells. In addition, an increased number of crypt cells harboring the oxidized DNA base 8-OHdG and $\gamma$H2AX foci were also detected in the intestines of OH-GQD-treated mice. Mechanistically, the OH-GQDs up-regulated both total and phosphorylated p53. Consistent with this, the average number of TUNEL+ and cleaved caspase-3+ apoptotic intestinal epithelial cells were significantly increased after OH-GQDs treatment. Finally, a 3-dimensional organoid culture was established using isolated crypts, and OH-GQDs treatment significantly reduced the size of the surviving intestinal organoids. Taken together, the intestinal toxicity of the OH-GQDs should be taken into account during biomedical applications.
IL-27 amplifies cytokine responses to Gram-negative bacterial products and Salmonella typhimurium infection. C. Petes et al. Scientific Reports 2018 SEP
Abstract
Cytokine responses from monocytes and macrophages exposed to bacteria are of particular importance in innate immunity. Focusing on the impact of the immunoregulatory cytokine interleukin (IL)-27 on control of innate immune system responses, we examined human immune responses to bacterial products and bacterial infection by E. coli and S. typhimurium. Since the effect of IL-27 treatment in human myeloid cells infected with bacteria is understudied, we treated human monocytes and macrophages with IL-27 and either LPS, flagellin, or bacteria, to investigate the effect on inflammatory signaling and cytokine responses. We determined that simultaneous stimulation with IL-27 and LPS derived from E. coli or S. typhimurium resulted in enhanced IL-12p40, TNF-$\alpha$, and IL-6 expression compared to that by LPS alone. To elucidate if IL-27 manipulated the cellular response to infection with bacteria, we infected IL-27 treated human macrophages with S. typhimurium. While IL-27 did not affect susceptibility to S. typhimurium infection or S. typhimurium-induced cell death, IL-27 significantly enhanced proinflammatory cytokine production in infected cells. Taken together, we highlight a role for IL-27 in modulating innate immune responses to bacterial infection.
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