Characterization of mouse lymphohematopoietic stem cells lacking spleen colony-forming activity.
The classical definition of lymphohematopoietic stem cells (LHSC),the most primitive progenitors of all blood cells,requires that they have the capacity for self-renewal and for the long-term production of all blood cell lineages. However,other characteristics of LHSC have been debated. Our previous data suggested that mouse LHSC are very slowly proliferating cells that generate delayed multilineage engraftment,while radioprotection" (rapid engraftment that will prevent early death from radiation-induced marrow aplasia) results from more committed progenitors. Alternatively�
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T. Derakhshan et al. ( 2018)
Stem cells international 2018 2136193
Development of Human Mast Cells from Hematopoietic Stem Cells within a 3D Collagen Matrix: Effect of Stem Cell Media on Mast Cell Generation.
Mast cells (MCs) arise from hematopoietic stem cells (HSCs) that mature within vascularized tissues. Fibroblasts and endothelial cells (ECs) play a role in the maturation of HSCs in the tissues. Due to difficulties in isolating MCs from tissues,large numbers of committed MC precursors can be generated in 2D culture systems with the use of differentiation factors. Since MCs are tissue-resident cells,the development of a 3D tissue-engineered model with ancillary cells that more closely mimics the 3D in vivo microenvironment has greater relevance for MC studies. The goals of this study were to show that MCs can be derived from HSCs within a 3D matrix and to determine a media to support MCs,fibroblasts,and ECs. The results show that HSCs within a collagen matrix cultured in StemSpan media with serum added at the last week yielded a greater number of c-kit+ cells and a greater amount of histamine granules compared to other media tested. Media supplemented with serum were necessary for EC survival,while fibroblasts survived irrespective of serum with higher cell yields in StemSpan. This work demonstrates the development of functional MCs within a 3D collagen matrix using a stem cell media that supports fibroblast and ECs.
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产品类型：
产品号#：
09600
09650
产品名：
StemSpan™ SFEM
StemSpan™ SFEM
Ibarra I et al. (DEC 2007)
Genes & development 21 24 3238--43
A role for microRNAs in maintenance of mouse mammary epithelial progenitor cells.
microRNA (miRNA) expression profiles are often characteristic of specific cell types. The mouse mammary epithelial cell line,Comma-Dbeta,contains a population of self-renewing progenitor cells that can reconstitute the mammary gland. We purified this population and determined its miRNA signature. Several microRNAs,including miR-205 and miR-22,are highly expressed in mammary progenitor cells,while others,including let-7 and miR-93,are depleted. Let-7 sensors can be used to prospectively enrich self-renewing populations,and enforced let-7 expression induces loss of self-renewing cells from mixed cultures.
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Eminli S et al. (SEP 2009)
Nature genetics 41 9 968--76
Differentiation stage determines potential of hematopoietic cells for reprogramming into induced pluripotent stem cells.
The reprogramming of somatic cells into induced pluripotent stem (iPS) cells upon overexpression of the transcription factors Oct4,Sox2,Klf4 and cMyc is inefficient. It has been assumed that the somatic differentiation state provides a barrier for efficient reprogramming; however,direct evidence for this notion is lacking. Here,we tested the potential of mouse hematopoietic cells at different stages of differentiation to be reprogrammed into iPS cells. We show that hematopoietic stem and progenitor cells give rise to iPS cells up to 300 times more efficiently than terminally differentiated B and T cells do,yielding reprogramming efficiencies of up to 28%. Our data provide evidence that the differentiation stage of the starting cell has a critical influence on the efficiency of reprogramming into iPS cells. Moreover,we identify hematopoietic progenitors as an attractive cell type for applications of iPS cell technology in research and therapy.
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产品类型：
产品号#：
02690
09600
09650
200-0000
200-0001
200-0002
70008
70008.1
70008.2
70008.3
70008.4
70008.5
85850
85857
产品名：
StemSpan™CC100
StemSpan™ SFEM
StemSpan™ SFEM
冻存的人脐带血CD34+细胞
冻存的人脐带血CD34+细胞
冻存的人脐带血CD34+细胞
冻存的人脐带血CD34+细胞
冻存的人脐带血CD34+细胞
冻存的人脐带血CD34+细胞
冻存的人脐带血CD34+细胞
冻存的人脐带血CD34+细胞
冻存的人脐带血CD34+细胞
mTeSR™1
mTeSR™1
Aguila JR et al. (JUL 2011)
Blood 118 3 576--85
SALL4 is a robust stimulator for the expansion of hematopoietic stem cells.
HSCs are rare cells that have the unique ability to self-renew and differentiate into cells of all hematopoietic lineages. The lack of donors and current inability to rapidly and efficiently expand HSCs are roadblocks in the development of successful cell therapies. Thus,the challenge of ex vivo human HSC expansion remains a fertile and critically important area of investigation. Here,we show that either SALL4A- or SALL4B-transduced human HSCs obtained from the mobilized peripheral blood are capable of rapid and efficient expansion ex vivo by textgreater10 000-fold for both CD34(+)/CD38(-) and CD34(+)/CD38(+) cells in the presence of appropriate cytokines. We found that these cells retained hematopoietic precursor cell immunophenotypes and morphology as well as normal in vitro or vivo potential for differentiation. The SALL4-mediated expansion was associated with enhanced stem cell engraftment and long-term repopulation capacity in vivo. Also,we demonstrated that constitutive expression of SALL4 inhibited granulocytic differentiation and permitted expansion of undifferentiated cells in 32D myeloid progenitors. Furthermore,a TAT-SALL4B fusion rapidly expanded CD34(+) cells,and it is thus feasible to translate this study into the clinical setting. Our findings provide a new avenue for investigating mechanisms of stem cell self-renewal and achieving clinically significant expansion of human HSCs.
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产品类型：
产品号#：
09600
09650
产品名：
StemSpan™ SFEM
StemSpan™ SFEM
Orelio C et al. (DEC 2008)
Blood 112 13 4895--904
Interleukin-1-mediated hematopoietic cell regulation in the aorta-gonad-mesonephros region of the mouse embryo.
Hematopoiesis during development is a dynamic process,with many factors involved in the emergence and regulation of hematopoietic stem cells (HSCs) and progenitor cells. Whereas previous studies have focused on developmental signaling and transcription factors in embryonic hematopoiesis,the role of well-known adult hematopoietic cytokines in the embryonic hematopoietic system has been largely unexplored. The cytokine interleukin-1 (IL-1),best known for its proinflammatory properties,has radioprotective effects on adult bone marrow HSCs,induces HSC mobilization,and increases HSC proliferation and/or differentiation. Here we examine IL-1 and its possible role in regulating hematopoiesis in the midgestation mouse embryo. We show that IL-1,IL-1 receptors (IL-1Rs),and signaling mediators are expressed in the aorta-gonad-mesonephros (AGM) region during the time when HSCs emerge in this site. IL-1 signaling is functional in the AGM,and the IL-1RI is expressed ventrally in the aortic subregion by some hematopoietic,endothelial,and mesenchymal cells. In vivo analyses of IL-1RI-deficient embryos show an increased myeloid differentiation,concomitant with a slight decrease in AGM HSC activity. Our results suggest that IL-1 is an important homeostatic regulator at the earliest time of HSC development,acting to limit the differentiation of some HSCs along the myeloid lineage.
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产品类型：
产品号#：
03434
03444
产品名：
MethoCult™GF M3434
MethoCult™GF M3434
J. W. Schott et al. (sep 2019)
Molecular therapy. Methods {\&} clinical development 14 134--147
Enhancing Lentiviral and Alpharetroviral Transduction of Human Hematopoietic Stem Cells for Clinical Application.
Ex vivo retroviral gene transfer into CD34+ hematopoietic stem and progenitor cells (HSPCs) has demonstrated remarkable clinical success in gene therapy for monogenic hematopoietic disorders. However,little attention has been paid to enhancement of culture and transduction conditions to achieve reliable effects across patient and disease contexts and to maximize potential vector usage and reduce treatment cost. We systematically tested three HSPC culture media manufactured to cGMP and eight previously described transduction enhancers (TEs) to develop a state-of-the-art clinically applicable protocol. Six TEs enhanced lentiviral (LV) and five TEs facilitated alpharetroviral (ARV) CD34+ HSPC transduction when used alone. Combinatorial TE application tested with LV vectors yielded more potent effects,with up to a 5.6-fold increase in total expression of a reporter gene and up to a 3.8-fold increase in VCN. Application of one of the most promising combinations,the poloxamer LentiBOOST and protamine sulfate,for GMP-compliant manufacturing of a clinical-grade advanced therapy medicinal product (ATMP) increased total VCN by over 6-fold,with no major changes in global gene expression profiles or inadvertent loss of CD34+CD90+ HSPC populations. Application of these defined culture and transduction conditions is likely to significantly improve ex vivo gene therapy manufacturing protocols for HSPCs and downstream clinical efficacy.
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产品类型：
产品号#：
19254
19254RF
产品名：
EasySep™人Naïve B细胞富集试剂盒
RoboSep™ 人Naïve B细胞富集试剂盒含滤芯吸头
C. T. Charlesworth et al. (SEP 2018)
Molecular therapy. Nucleic acids 12 89--104
Priming Human Repopulating Hematopoietic Stem and Progenitor Cells for Cas9/sgRNA Gene Targeting.
Engineered nuclease-mediated gene targeting through homologous recombination (HR) in hematopoietic stem and progenitor cells (HSPCs) has the potential to treat a variety of genetic hematologic and immunologic disorders. Here,we identify critical parameters to reproducibly achieve high frequencies of RNA-guided (single-guide RNA [sgRNA]; CRISPR)-Cas9 nuclease (Cas9/sgRNA) and rAAV6-mediated HR at the $\beta$-globin (HBB) locus in HSPCs. We identified that by transducing HSPCs with rAAV6 post-electroporation,there was a greater than 2-fold electroporation-aided transduction (EAT) of rAAV6 endocytosis with roughly 70{\%} of the cell population having undergone transduction within 2 hr. When HSPCs are cultured at low densities (1 × 105 cells/mL) prior to HBB targeting,HSPC expansion rates are significantly positively correlated with HR frequencies in vitro as well as in repopulating cells in immunodeficient NSG mice in vivo. We also show that culturing fluorescence-activated cell sorting (FACS)-enriched HBB-targeted HSPCs at low cell densities in the presence of the small molecules,UM171 and SR1,stimulates the expansion of gene-edited HSPCs as measured by higher engraftment levels in immunodeficient mice. This work serves not only as an optimized protocol for genome editing HSPCs at the HBB locus for the treatment of $\beta$-hemoglobinopathies but also as a foundation for editing HSPCs at other loci for both basic and translational research.
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EasySep™小鼠TIL（CD45）正选试剂盒
50:59
线上讲座Gene Targeting in Hematopoietic Stem Cells for Basic and Translational Research发布日期: 11/28/2018
科学海报Improved Ex Vivo Expansion of Hematopoietic Progenitor Cells from Cord Blood in a Novel Serum and Animal Component Free Medium
科学海报Optimized Media and Workflow for the Expansion of Human Pluripotent Stem Cells as Aggregates in Suspension Cultures
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