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Figure 1. Typical EasySep™ Human CD34 Positive Selection Kit II Isolation Profile
Starting with cord blood, mobilized peripheral blood or bone marrow MNCs, or ES and iPS cell cultures, the CD34+ cell content of the isolated fraction is typically 93.5 ± 1.1% (mean ± SD), using the purple EasySep™ Magnet. In the above example using frozen cord blood, the purities of the start and final isolated fractions are 2.2% and 94.7%, respectively.
Figure 2. FACS Data for Anti-Human CD34 Antibody, Clone 581, Alexa Fluor® 488-Conjugated
(A) Flow cytometry analysis of human peripheral blood mononuclear cells (PBMCs) labeled with Anti-Human CD34 Antibody, Clone 581, Alexa Fluor® 488 (Catalog #60013AD) and Anti-Human CD45 Antibody, Clone HI30, APC (Catalog #60018AZ). (B) Flow cytometry analysis of PBMCs labeled with Mouse IgG1, kappa Isotype Control Antibody, Clone MOPC-21, Alexa Fluor® 488 (Catalog #60070AD), and Anti-Human CD45 Antibody, Clone HI30, APC. (C) Flow cytometry analysis of human PBMCs processed with the EasySep™ Human CD34 Positive Selection Kit (Catalog #17856) and labeled with Anti-Human CD34 Antibody, Clone 581, APC. Histograms show labeling of PBMCs (Start) and isolated cells (Isolated). Labeling of start cells with Mouse IgG1, kappa Isotype Control Antibody, Clone MOPC-21, Alexa Fluor® 488 is shown (solid line histogram).
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.
Identification of key microRNAs as predictive biomarkers of Nilotinib response in chronic myeloid leukemia: a sub-analysis of the ENESTxtnd clinical trial. R. Yen et al. Leukemia 2022 oct
Abstract
Despite the effectiveness of tyrosine kinase inhibitors (TKIs) against chronic myeloid leukemia (CML), they are not usually curative as some patients develop drug-resistance or are at risk of disease relapse when treatment is discontinued. Studies have demonstrated that primitive CML cells display unique miRNA profiles in response to TKI treatment. However, the utility of miRNAs in predicting treatment response is not yet conclusive. Here, we analyzed differentially expressed miRNAs in CD34+ CML cells pre- and post-nilotinib (NL) therapy from 58 patients enrolled in the Canadian sub-analysis of the ENESTxtnd phase IIIb clinical trial which correlated with sensitivity of CD34+ cells to NL treatment in in vitro colony-forming cell (CFC) assays. We performed Cox Proportional Hazard (CoxPH) analysis and applied machine learning algorithms to generate multivariate miRNA panels which can predict NL response at treatment-na{\{i}}ve or post-treatment time points. We demonstrated that a combination of miR-145 and miR-708 are effective predictors of NL response in treatment-na{\"{i}}ve patients whereas miR-150 and miR-185 were significant classifiers at 1-month and 3-month post-NL therapy. Interestingly incorporation of NL-CFC output in these panels enhanced predictive performance. Thus this novel predictive model may be developed into a prognostic tool for use in the clinic."
Enhancement of proliferation of human umbilical cord blood-derived CD34+ hematopoietic stem cells by a combination of hyper-interleukin-6 and small molecules. Y. S. Park et al. Biochemistry and biophysics reports 2022 mar
Abstract
Umbilical cord blood (UCB) is an alternative source of allogeneic hematopoietic stem cells (HSCs) for transplantation to treat various hematological disorders. The major limitation to the use of UCB-derived HSCs (UCB-HSCs) in transplantation, however, is the low numbers of HSCs in a unit of cord blood. To overcome this limitation, various cytokines or small molecules have been used to expand UCB-HSCs ex vivo. In this study, we investigated a synergistic effect of the combination of HIL-6, SR1, and UM171 on UCB-HSC culture and found that this combination resulted in the highest number of CD34+ cells. These results suggest that the combination of SR1, UM171 and HIL-6 exerts a synergistic effect in the proliferation of HSCs from UCB and thus, SR1, UM171 and HIL-6 is the most suitable combination for obtaining HSCs from UCB for clinical transplantation.
Asymmetric organelle inheritance predicts human blood stem cell fate. D. Loeffler et al. Blood 2022 mar
Abstract
Understanding human hematopoietic stem cell fate control is important for its improved therapeutic manipulation. Asymmetric cell division, the asymmetric inheritance of factors during division instructing future daughter cell fates, was recently described in mouse blood stem cells. In human blood stem cells, the possible existence of asymmetric cell division remained unclear because of technical challenges in its direct observation. Here, we use long-term quantitative single-cell imaging to show that lysosomes and active mitochondria are asymmetrically inherited in human blood stem cells and that their inheritance is a coordinated, nonrandom process. Furthermore, multiple additional organelles, including autophagosomes, mitophagosomes, autolysosomes, and recycling endosomes, show preferential asymmetric cosegregation with lysosomes. Importantly, asymmetric lysosomal inheritance predicts future asymmetric daughter cell-cycle length, differentiation, and stem cell marker expression, whereas asymmetric inheritance of active mitochondria correlates with daughter metabolic activity. Hence, human hematopoietic stem cell fates are regulated by asymmetric cell division, with both mechanistic evolutionary conservation and differences to the mouse system.
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