hematopoietic stem cell donors  (CH Instruments)

Journal: Journal for Immunotherapy of Cancer

Article Title: T cell engaging bispecific antibodies targeting CD33 IgV and IgC domains for the treatment of acute myeloid leukemia

doi: 10.1136/jitc-2021-002509

Figure Lengend Snippet: BC275 does not bind to hematopoietic stem and progenitor cells. Freshly isolated T cells (A) and peripheral blood mononuclear cells (PBMCs) (B) were cultured without target cells or with CD33(+) acute myeloid leukemia (AML) line Kasumi-1 in the presence of dilutions of the CD33xCD3 bispecific antibodies (BsAbs) for 1 day with an E:T ratio of 10. The production of Th1 cytokines was measured by a bead-based assay and flow cytometry. The binding of the BsAbs to bone marrow (BM) or cord blood (CB) CD34(+) hematopoietic stem and progenitor cells was assessed by flow cytometry (C). CD33(+) AML cell lines Kasumi-1, MOLM13, and SKM1 were used as positive controls. IFN-γ, interferon-γ; IL, interleukin; TNF-α, tumor necrosis factor-α.

Article Snippet: Cryopreserved bone marrow (BM) and cord blood (CB)-derived CD34(+) hematopoietic stem and progenitor cells (HSPCs) were purchased from ZenBio (Cat # SER-BMCD34-F and SER-CD34-1F, respectively).

Techniques: Isolation, Cell Culture, Bead-based Assay, Flow Cytometry, Binding Assay

Journal: Experimental hematology

Article Title: Compounds targeting Class II histone deacetylases do not cause panHDACi associated impairment of megakaryocyte differentiation

doi: 10.1016/j.exphem.2018.12.007

Figure Lengend Snippet: CD34+ HSC cells were cultured and induced to differentiate to MK in the presence of 0.1% DMSO only (control), LMK235 (75 and 200 nM), Panobinostat (5 nM), or SAHA (500 nM) for 14-days followed by FACS analyses.

Article Snippet: Human CD34 + hematopoietic stem cells (mixed donor) were purchased from Stem Cell Technologies (cat#70008, Vancouver, Canada) and expanded in StemSpan SFEMII media (cat# 09605) supplemented with StemSpan megakaryocytes (MK) expansion supplement (cat# 02696; both from Stem Cell Technologies) to induce differentiation.

Techniques: Cell Culture, Control

Journal: Experimental hematology

Article Title: Compounds targeting Class II histone deacetylases do not cause panHDACi associated impairment of megakaryocyte differentiation

doi: 10.1016/j.exphem.2018.12.007

Figure Lengend Snippet: A. HSC CD34+ cells were plated at 30,000 cells/mL and were induced to differentiation for 8 days in the presence of 0.1% DMSO (control), LMK235 or SAHA and Panobinostat as positive controls. Data indicate that LMK235 at 75, 200 and 400 nM resulted in the cell number and morphology of MK similar to those of DMSO treated control cells. Panobinostat treatment resulted in a significant reduction in the cell number and morphologically smaller cells, indicating immature cells. Similarly, SAHA (at 500 nM) reduced the overall number of cells, however, some large MK were evident. Images were captured using 40 × objective.

Article Snippet: Human CD34 + hematopoietic stem cells (mixed donor) were purchased from Stem Cell Technologies (cat#70008, Vancouver, Canada) and expanded in StemSpan SFEMII media (cat# 09605) supplemented with StemSpan megakaryocytes (MK) expansion supplement (cat# 02696; both from Stem Cell Technologies) to induce differentiation.

Techniques: Control

Journal: Experimental hematology

Article Title: Compounds targeting Class II histone deacetylases do not cause panHDACi associated impairment of megakaryocyte differentiation

doi: 10.1016/j.exphem.2018.12.007

Figure Lengend Snippet: A. Primary CD34+ cells were induced to differentiate to MK cells for 14 days in the presence of 0.1% DMSO (control) or tubacin at 300 nM. Flow cytometric analysis was carried out subsequently. Top row: Dot plot of forward scatter (FSC) versus side scatter (SSC) outlining MK cell size and shape, respectively. Two distinct populations (P1 and P2) of high scatter cells, were identified and further characterized for MK cell maturity. Middle Row: To evaluate MK cell maturity, cells were labeled with FITC anti-CD41 (marker present on all MK cells) and PE-anti-CD42b (mature MK cell marker). Analytical gate was set to include only viable cells (7-AAD negative). Dot plot upper right quadrant (Q2) indicates mature, differentiated MK cells. As part of the high scatter population characterization, P1 cell population was labeled red, higher FSC population P2 a blue color and the rest of the viable cell were black. Bottom row: Cell ploidy was evaluated by staining DNA with propidium iodide (PI) and quantifying PI ploidy classes in CD41+ cells. The arrows indicate ploidy classes from 2N to >32N. All experiments were performed three times and representative results were shown.

Article Snippet: Human CD34 + hematopoietic stem cells (mixed donor) were purchased from Stem Cell Technologies (cat#70008, Vancouver, Canada) and expanded in StemSpan SFEMII media (cat# 09605) supplemented with StemSpan megakaryocytes (MK) expansion supplement (cat# 02696; both from Stem Cell Technologies) to induce differentiation.

Techniques: Control, Labeling, Marker, Staining

Journal: Experimental hematology

Article Title: Compounds targeting Class II histone deacetylases do not cause panHDACi associated impairment of megakaryocyte differentiation

doi: 10.1016/j.exphem.2018.12.007

Figure Lengend Snippet: Based on current and published literature, we conclude that panHDACi cause impairment of megakaryopoiesis mainly through the inhibition of HDAC1 and HDAC2. Since HDAC1,2 function is required for GATA-1 stabilization, the inhibition of HDAC1,2 leads to GATA-1 degradation, which may be mediated by GATA-1 acetylation. Compounds specifically targeting Class II HDACs such as 4, 5 and 6 avoid disrupting HDAC1,2 function, thus avoiding GATA-1 degradation and subsequent impairment of differentiation of hematopoietic progenitor cells. Therefore, specifically targeting class II HDACs as part of cancer therapy, may emerge as a safer approach than non-selective HDAC inhibition.

Article Snippet: Human CD34 + hematopoietic stem cells (mixed donor) were purchased from Stem Cell Technologies (cat#70008, Vancouver, Canada) and expanded in StemSpan SFEMII media (cat# 09605) supplemented with StemSpan megakaryocytes (MK) expansion supplement (cat# 02696; both from Stem Cell Technologies) to induce differentiation.

Techniques: Inhibition