Journal: bioRxiv

Article Title: Development of Scaffold-free 3D Cholangiocyte Organoids to Study the Progression of Primary Sclerosing Cholangitis

doi: 10.1101/2022.10.24.513594

Figure Lengend Snippet: A . IF staining for Desmin (red), and αSMA (green) of immortalized human HSCs from normal and PSC patients. Right, quantification of Desmin + and αSMA + cell percentage (% Hoechst stained nucleus) in isolated normal and PSC HSCs (n=3 patients for normal and PSC) (* = p <0.05). B . IF staining for GFAP (red), and POSTN (green) of immortalized human HSCs from normal and PSC patients. Right, quantification of GFAP + and POSTN + cells (% Hoechst stained nucleus) in isolated normal and PSC HSCs (n=3 patients for normal and PSC) (* = p <0.05). C . IF staining for CD31 (green) of immortalized human LECs from normal and PSC patients. Hoechst (blue) staining for nucleus counterstain. Right, quantification of CD31 + cells (% Hoechst stained nucleus) in isolated normal and PSC LECs. Scale bars: 100µm. D . qRT-PCR analysis of the expression of Desmin and PECAM in HSCs and LECs from normal and PSC patients, confirming cell phenotypes and characteristics.

Article Snippet: The NPCs that remained in the supernatant were centrifuged at 600x g for 10 minutes at 4°C, and then resuspended to purify LECs with FACS to select CD31 + cells (Miltenyi Biotech, Bergisch Gladbach, Germany, Catalog No. Cat# 130-117-312).

Techniques: Staining, Isolation, Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: Development of Scaffold-free 3D Cholangiocyte Organoids to Study the Progression of Primary Sclerosing Cholangitis

doi: 10.1101/2022.10.24.513594

Figure Lengend Snippet: A . Representative brightfield images of 3-cell 3D-CHOs (cholangiocytes, LECs, HSCs) from normal and PSC patients on day 1 and day 4. A large number of cell aggregates (within the inner white line circle and outer layer circle) detached from the main organoid (within the inner circle) were observed in PSC organoids on day 4. B . CellTiterGlo 3D assay on normal and PSC 3D-CHOs showing significant proliferation in PSC cells after culturing for 5 days (n=3 per normal and PSC organoids). C . Top: representative IF co-staining image for CK-19 (in red for cholangiocytes) and αSMA (in green for HSCs) in normal and PSC organoids. Middle: representative IF co-staining image for CFTR (in green for cholangiocytes) and COL1A1 (in red for HSCs) in normal and PSC organoids. The side magnified image shows a nodular-like image in PSC organoids. Bottom: representative IF staining image for CD31 (in green for LEC vascularization) in normal and PSC organoids. Cell nucleus was stained with Hoechst (in blue). Scale bars: 100µm. D . qRT-PCRs for cholangiocyte marker genes Sox9, EpCAM, SCT (secretin) and SCTR (secretin receptor), fibrosis marker genes ACTA2, COL1A1, DESMIN (DES), and TGFβ1, angiogenesis marker genes PECAM, CDH5, and vWF, and inflammation marker genes IL-6 and TNFα. *P<0.05, **p<0.01 PSC vs. normal organoids. (n=4 organoids per each experiment).

Article Snippet: The NPCs that remained in the supernatant were centrifuged at 600x g for 10 minutes at 4°C, and then resuspended to purify LECs with FACS to select CD31 + cells (Miltenyi Biotech, Bergisch Gladbach, Germany, Catalog No. Cat# 130-117-312).

Techniques: Staining, Marker

Journal: iScience

Article Title: In vitro vascularized immunocompetent patient-derived model to test cancer therapies

doi: 10.1016/j.isci.2023.108094

Figure Lengend Snippet:

Article Snippet: Mouse anti-CD31 , Cell Applications , Cat# CB13678.

Techniques: Plasmid Preparation, Recombinant, Lysis, RNA Extraction, Amplification, DC Protein Assay, Enzyme-linked Immunosorbent Assay, Sequencing, Software, Fluorescence, Microscopy

Journal: Drug Design, Development and Therapy

Article Title: MK-4 Ameliorates Diabetic Osteoporosis in Angiogenesis-Dependent Bone Formation by Promoting Mitophagy in Endothelial Cells

doi: 10.2147/DDDT.S503930

Figure Lengend Snippet: MK-4 ameliorates the impairments of type H vessel formation and angiogenesis-dependent bone formation in DOP mice. ( a ) Schematic drawing of the procedures conducted in this study. ( b and c ) Representative flow cytometry plots of CD31 hi Emcn hi ECs (type H ECs). The bar graph showed the quantitation of type H ECs. ( d and e ) Confocal image of CD31 hi and Emcn hi (type H blood vessels) in mice models and histomorphometric quantitation of the area of type H blood vessels. Scale bar: 50μm. ( f and g ) Confocal image of Osterix + (green) osteoprogenitors around CD31 hi vessels (red) and the quantitative analysis. Scale bar: 50μm. ( h ) Typical three-dimensional and two-dimensional coronal images of the tibias were obtained by Micro-CT. ( i ) HE and Masson staining of mice tibias. Scale bar: 200μm or 100μm. ( j ) Statistical analysis of bone histological parameters. ( k and l ) IHC analysis of ALP and RUNX2 in the tibias of mice models. Scale bar: 50μm. Data were presented as mean ± SD (n=5). * P < 0.05. ** P < 0.01. *** P < 0.001.

Article Snippet: The cells were subsequently washed and later incubated with PE-conjugated CD45 (E-AB-F1136D, 1:20; Elabscience, Wuhan, China) and APC-conjugated CD31 (E-AB-F1180E, 1:20; Elabscience) antibodies at 4 °C for 45 minutes.

Techniques: Flow Cytometry, Quantitation Assay, Micro-CT, Staining

Journal: Drug Design, Development and Therapy

Article Title: MK-4 Ameliorates Diabetic Osteoporosis in Angiogenesis-Dependent Bone Formation by Promoting Mitophagy in Endothelial Cells

doi: 10.2147/DDDT.S503930

Figure Lengend Snippet: 3-MA partially counteracts the promotion of type H vessel recovery and angiogenesis-dependent bone formation gain by MK-4. ( a and b ) Confocal image of CD31 hi and Emcn hi (termed type H blood vessels) in mice models and histomorphometric quantitation of the area of type H blood vessels. Scale bar: 50μm. ( c and d ) Confocal image of Osterix + (red) osteoprogenitors around Emcn hi vessels (green) and the quantitative analysis. Scale bar: 50μm. ( e ) HE and Masson staining of mice tibias. Scale bar: 200μm or 100μm. ( f ) Statistical analysis of bone histological parameters. ( g and h ) IHC analysis of ALP and RUNX2 in the tibias of mice models. Scale bar: 50μm. Data were presented as mean ± SD (n=5). * P < 0.05. ** P < 0.01. *** P < 0.001.

Article Snippet: The cells were subsequently washed and later incubated with PE-conjugated CD45 (E-AB-F1136D, 1:20; Elabscience, Wuhan, China) and APC-conjugated CD31 (E-AB-F1180E, 1:20; Elabscience) antibodies at 4 °C for 45 minutes.

Techniques: Quantitation Assay, Staining