Journal: Cell Proliferation

Article Title: CD34 dim cells identified as pluripotent stem cell‐derived definitive hemogenic endothelium purified using bone morphogenetic protein 4

doi: 10.1111/cpr.13366

Figure Lengend Snippet: Purified hemogenic endothelium (HE) can differentiate into all haematopoietic lineage cells. (A) Cell morphology in HE before and after CD34 sorting. Purified and homogenous HE underwent EHT on Day 8 after sorting. Magnification, 20×. (B) CD34 purification was confirmed and mRNA expression of representative haematopoietic lineage committed factors in CD34 + and CD34 − cells measured by qRT‐PCR. (C) Giemsa staining in differentiated cells from purified HE. Scale bar, 20 μM. All haematopoietic lineage cells were generated from CD34‐sorted HE within 32 days. Lineage cell differentiation from HE was confirmed using specific lineage markers. Black arrows, shedding platelets, MK‐III types. Magnification, 20×. (D) Flow cytometry data show abundant haematopoietic progenitors, implying full commitment toward blood lineages. CD45 + CD34 + CD3 + cells also emerged from purified HE from Day 10 ( n = 4–6). Markers of maturation also increased in differentiated cells by Day 40. Data are derived from at least two independent experiments with duplicates ( n = 4–7). *** p < 0.001

Article Snippet: Hemogenic ECs derived from PSCs were dropped onto microscope slides and air‐dried, then haematopoietic lineage cells were stained with Wright‐Giemsa Solution (ab245888, Abcam) as described by the manufacturer.

Techniques: Purification, Expressing, Quantitative RT-PCR, Staining, Generated, Cell Differentiation, Flow Cytometry, Derivative Assay

Journal: Cells

Article Title: NAD Modulates DNA Methylation and Cell Differentiation

doi: 10.3390/cells10112986

Figure Lengend Snippet: NAD induces myeloid differentiation in K562. ( a ) Wright Giemsa staining showing morphological changes between NAD-treated and control cells after four days. The figure is representative of five independent experiments ( b ) Representative image showing the increase in the surface markers CD15, CD14 and CD11b upon NAD treatment by flow cytometry. The table reports the mean % ±SD of CD14 + , CD15 + , CD11b + cells of five independent experiments. *: p < 0.05; **: p < 0.01; ***: p < 0.001 vs. vehicle ( n = 5) ( c ) NBT positive staining detected by small blue dots after counterstaining the cells with safranin. A magnification is shown in the rectangle. The pictures are representative of five independent experiments ( d ) Seahorse XF analysis of K562 mitochondrial stress response in cells treated with NAD or vehicle. The figure represents the mean of three biological replicates ( n = 3). Error bars indicate ± s.d.

Article Snippet: The cells were then stained with the Wright Giemsa solutions kit (CAMCO STAIN PAK Fort Lauderdale, FL, USA, pc#702) according to manufacturer’s instructions.

Techniques: Staining, Control, Flow Cytometry