easysep human cord blood cd34 positive selection kit ii  (STEMCELL Technologies Inc)

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Comparison of NSG-Quad and MISTRG-6 humanized mice for modeling circulating and tumor-infiltrating human myeloid cells

doi: 10.1016/j.omtm.2025.101487

Figure Lengend Snippet: NSG-Quad mice support human hematopoietic cell engraftment and multilineage immune cell development (A) Percentage of human CD45 + (hCD45 + ) cells of total CD45 + cells (mouse and human) in the blood at 8 weeks and 10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells. (B) Human immune cell composition in the blood of NSG ( n = 33), NSG-Quad ( n = 34), and MISTRG-6 mice ( n = 32) 8 weeks post-engraftment and in the blood of NSG ( n = 29), NSG-Quad ( n = 25), and MISTRG-6 mice ( n = 16) 10–15 weeks post-engraftment. B cells (CD3 − CD56 − CD33 − CD20 + ), myeloid cells (CD3 − CD56 − CD20 − CD33 + ), NK cells (CD3 − CD56 + ), CD8 + T cells (CD3 + CD4 − CD8 + ), and CD4 + T cells (CD3 + CD8 − CD4 + ). (C) Percentage of human immune cell subsets in the blood 8 weeks post-engraftment (data from B). (D) Percentage of human immune cell subsets in the blood 10–15 weeks post-engraftment (data from B). Data are shown as mean ± SEM. p values were calculated using one-way or two-way ANOVA (A) with Tukey’s multiple comparison test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Human CD34 + HSPCs were isolated from cord blood using an EasySep Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies, #17896).

Techniques: Derivative Assay, Comparison

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Comparison of NSG-Quad and MISTRG-6 humanized mice for modeling circulating and tumor-infiltrating human myeloid cells

doi: 10.1016/j.omtm.2025.101487

Figure Lengend Snippet: NSG-Quad mice support the development of human myeloid cells (A) Percentage of human CD45 + (hCD45 + ) cells of total CD45 + cells (mouse and human) in the blood of NSG-Quad +/− mice (heterozygous M-CSF, n = 24) and NSG-Quad +/+ mice (homozygous M-CSF, n = 9) at 10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells. (B) Human immune cell composition in the blood of NSG-Quad +/− mice ( n = 24) and NSG-Quad +/+ mice ( n = 9) 10–15 weeks post-engraftment. (C) Percentage of human immune cell subsets in the blood of NSG-Quad +/− and NSG-Quad +/+ mice 10–15 weeks post-engraftment (data from B). (D) Percentage of human CD45 + (hCD45 + ) cells of total CD45 + cells (mouse and human) in the blood at 8 weeks and 10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells. (E) Human immune cell composition in the blood of NSGS ( n = 11) and NSG-Quad mice ( n = 25) 10–15 weeks post-engraftment. (F) Percentage of human immune cell subsets in the blood of NSGS and NSG-Quad mice 10–15 weeks post-engraftment (data from E). Data are shown as mean ± SEM. p values were calculated using two-tailed, unpaired Student’s t test (A), two-way ANOVA with Tukey’s multiple comparison test (D) and two-tailed, unpaired Mann-Whitney U test (C, F). ∗ p < 0.05, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Human CD34 + HSPCs were isolated from cord blood using an EasySep Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies, #17896).

Techniques: Derivative Assay, Two Tailed Test, Comparison, MANN-WHITNEY

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Comparison of NSG-Quad and MISTRG-6 humanized mice for modeling circulating and tumor-infiltrating human myeloid cells

doi: 10.1016/j.omtm.2025.101487

Figure Lengend Snippet: NSG-Quad mice promote the development of tumor-infiltrating human macrophages (A) Tumor growth curves in NSG ( n = 9), NSG-Quad ( n = 12), and MISTRG-6 mice ( n = 15) engrafted with SW480 CRC cells and in NSG ( n = 10), NSG-Quad ( n = 4), and MISTRG-6 mice ( n = 14) engrafted with HCT116 CRC cells. (B) Frequency of hCD45 + cells in the tumor of NSG, NSG-Quad, and MISTRG-6 humanized mice engrafted with SW480 or HCT116 CRC cells (10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells; end of experiment). (C) Human immune cell composition in the blood of NSG, NSG-Quad, and MISTRG-6 humanized mice engrafted with SW480 or HCT116 CRC cells (10–15 weeks post-engraftment with human-cord-blood-derived CD34 + cells; end of experiment; data from G). (D) Human immune cell composition in the tumor xenografts of NSG ( n = 9, SW480; n = 10, HCT116), NSG-Quad ( n = 10, SW480; n = 3, HCT116), and MISTRG-6 mice ( n = 15, SW480; n = 12, HCT116). (E) Representative IHC pictures show SW480 CRC-infiltrating human CD68 + macrophages in NSG, NSG-Quad, and MISTRG-6 humanized mice. Scale bar: 50 μm. (F) Frequency of human CD68 + macrophages in CRC xenografts of NSG ( n = 11), NSG-Quad ( n = 7), and MISTRG-6 mice ( n = 15) analyzed by IHC. (G) Frequency of CD86 + (M1-like) and CD163 + (M2-like) human CD14 + monocytes in SW480 and HCT116 CRC xenografts of NSG-Quad ( n = 13) and MISTRG-6 mice ( n = 17). (H) Composition of the human CD33 + CD66b − myeloid cell population based on the expression of CD14, CD16, and HLA-DR in SW480 CRC xenografts of NSG-Quad ( n = 8) and MISTRG-6 ( n = 3) mice. Data are shown as mean ± SEM. p values were calculated using one-way or two-way ANOVA (G) with Tukey’s multiple comparison test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Human CD34 + HSPCs were isolated from cord blood using an EasySep Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies, #17896).

Techniques: Derivative Assay, Expressing, Comparison

Journal: Biomaterials

Article Title: Organ-on-a-chip model of vascularized human bone marrow niches

doi: 10.1016/j.biomaterials.2021.121245

Figure Lengend Snippet: (A) Marker expression of independently cultured monolayers of hFOB and two BMSC donors as determined by flow cytometry, n = 4. (B) Cartoon schematic of experimental design. Cord blood-derived endothelial cells (EC) seeded with either hFOB (OB, Endosteal Niche) or BMSC (Perivascular Niche) form microvascular networks in a period of 4–7 days. (C) BMoaC cultured with CellTracker™ Violet hFOB and Deep Red BMSC demonstrated stromal separation by niche. (D) Relative fluorescent area (RFA) of Violet hFOB in the endosteal niche compared to unstained BMSC within the perivascular niche and RFA of unstained hFOB in the endosteal niche to Deep Red BMSC in the perivascular niche. Fluorescent area was determined by normalizing to an unstained control. *p < 0.05, ***p < 0.001 by two-way ANOVA. (E) Osteopontin (green) was expressed in the (i) endosteal and (ii) perivascular niches. EC (red) counterstained with CD34. (F) SDF-1 was detected adjacent to EC (red) in the (i) endosteal niche with enhanced expression of SDF-1 in the (ii) perivascular niche. (G) EC (red) were adjacent to stromal cells that express leptin as seen in the (i) endosteal and (ii) perivascular niche. Nuclei counterstained with DAPI (blue). Scale bars: 500 μm and 50 μm for low and high magnification images, respectively.

Article Snippet: HSPC (CD34 + ) were isolated from cord blood using the EasySep TM Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies) according to the manufacturer’s guidelines.

Techniques: Marker, Expressing, Cell Culture, Flow Cytometry, Derivative Assay, Control

Journal: Biomaterials

Article Title: Organ-on-a-chip model of vascularized human bone marrow niches

doi: 10.1016/j.biomaterials.2021.121245

Figure Lengend Snippet: (A)–(F) Endothelial cell (red, CD31 or CD34) (A) laminin in the basement membrane, (B) SCF, (C) I-CAM, (D) V-CAM, (E) E-Selectin/CD62E and (F) Nestin. ROIs show z-stacks in either the (i) endosteal or (ii) perivascular niche. Nuclei were counterstained with DAPI (Blue). Scale bars: 500 μm and 50 μm for low and high magnification views, respectively. (G) BMoaC microvascular networks were i) perfused with 70 kDA TRITC-dextran and ii) had permeabilities of 16.96 × 10−7 cm/s and 5.19 × 10−7 cm/s for the endosteal (n = 3) and perivascular niches (n = 5), respectively.

Article Snippet: HSPC (CD34 + ) were isolated from cord blood using the EasySep TM Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies) according to the manufacturer’s guidelines.

Techniques: Membrane

Journal: Biomaterials

Article Title: Organ-on-a-chip model of vascularized human bone marrow niches

doi: 10.1016/j.biomaterials.2021.121245

Figure Lengend Snippet: (A) Experimental design. (B) (i) CD34+ cells isolated from human cord blood were 90.4% ± 6.37 CD34+ and 78.4% ± 8.57 CD34+/CD133+, n = 14. (ii) Approximately 40–60 CD34+ HSPC were loaded into each device, n = 21 devices. (C) BMoaC supported CD34+ blood vessels. (i,ii) Small, round CD34+ HSPC (arrows) were present along with Lin+ cells. Scale bars: 500 μm and 50 μm for low and high magnification images, respectively. (D) Cells isolated after 7, 10, and 14 days of culture in the BMoaC were analyzed for CD34/CD133 from both the endosteal and perivascular niches, biological replicates with n = 5. (E) Cells isolated after 7, 10, and 14 days of culture from either niche in the BMoaC consistently generated CFU-GEMM, biological replicates with n = 10–11. (F) Cells isolated after 10 and 14 days of culture within single niche (SN, endosteal or perivascular) controls produced CFU-GEMM; monolayer controls did not.

Article Snippet: HSPC (CD34 + ) were isolated from cord blood using the EasySep TM Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies) according to the manufacturer’s guidelines.

Techniques: Isolation, Generated, Produced

Journal: Biomaterials

Article Title: Organ-on-a-chip model of vascularized human bone marrow niches

doi: 10.1016/j.biomaterials.2021.121245

Figure Lengend Snippet: (A) After 10 days of culture, BMoaC contain cells expressing CD14 (Green) in both sides of the device and in the (i, ii) lumen of CD34+ blood vessels (red). Nuclei were counterstained with DAPI (blue). Scale bars: 500 μm and 50 μm for low and high magnification images, respectively. Cells that egressed from the bone marrow niche and into the adjacent fluidic line of devices were collected on Day 14 and analyzed by flow cytometry for (B) the leukocyte common antigen CD45 and (C) the myeloid marker CD33 and neutrophil marker CD66b. (D) BMoaC exposed to 0.5 M doxorubicin between days 8 and 10 of a 14 day culture produce significantly fewer egressed CD66b+/CD33− cells from the perivascular niche compared to control devices. **p < 0.01 by 2-way ANOVA followed by Sidak’s post hoc-analysis. (E) BMoaC exposed to 30 ng/mL G-CSF produced significantly more egressed CD66b+/CD33− cells from the perivascular niche compared to control devices. *p < 0.05, **p < 0.01 by 2-way ANOVA followed by Tukey’s post hoc-analysis.

Article Snippet: HSPC (CD34 + ) were isolated from cord blood using the EasySep TM Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies) according to the manufacturer’s guidelines.

Techniques: Expressing, Flow Cytometry, Marker, Control, Produced

Journal: Biomaterials

Article Title: Organ-on-a-chip model of vascularized human bone marrow niches

doi: 10.1016/j.biomaterials.2021.121245

Figure Lengend Snippet: (A) Experimental design. Breast cancer cells were loaded into the bottom chamber of the BMoaC on Day 4 and cultured for an additional 8 days. (B) The triple negative breast cancer cell line MDA-MB-231 (green) proliferated over time and migrated into the bottom portion of both the endosteal and perivascular niches. EC and HSPC (red) were labeled with CD34 antibody and nuclei (blue) were counterstained with DAPI. Scale bar: 500 μm. The fluorescent area of cancer cells in each chamber of the device was quantified. (C) MDA-MB-231 cells migrated towards the endosteal and perivascular niches after 8 days of culture in the BMoaC. (D) MDA-MB-231 cancer cells in the BMoaC perivascular and endosteal niches versus fibrin only controls expressed significantly more Ki-67, suggesting active proliferation. Cancer cells in the BMoaC perivascular niche expressed more Ki-67 than those in the bottom chamber. *p < 0.05, **p < 0.01, ***p < 0.001 by 2-way ANOVA followed by Tukey’s post hoc-analysis.

Article Snippet: HSPC (CD34 + ) were isolated from cord blood using the EasySep TM Human Cord Blood CD34 Positive Selection Kit II (StemCell Technologies) according to the manufacturer’s guidelines.

Techniques: Cell Culture, Labeling