Journal: Journal of Cellular and Molecular Medicine

Article Title: Endothelial cell differentiation of human breast tumour stem/progenitor cells

doi: 10.1111/j.1582-4934.2008.00338.x

Figure Lengend Snippet: Mammosphere generation, characterization and epithelial differentiation. Mammospheres were obtained by culture of dissociated cells from a human breast tumour in mammosphere medium containing EGF and bFGF. ( A ) Representative micrograph showing the ematoxilin and eosin staining of paraffin-embedded, formalin-fixed mammospheres. ( B ) Mammosphere clone formation analysis in secondary and tertiary passages. The number of mammosphere clones/100 cells generated from single cells increased in the third passage. ( C ) Representative FACS analysis of mammosphere cell clones showing the expression of CD44 and not CD24 and of VEGF receptor 1 (VEGFR1; the dark area indicates the specific Ab, the white area the isotypic control). ( D ) Representative immunofluorescence expression by mammosphere cell clones of the stem cell markers Oct-4 and nestin, but not of the differentiation markers CK14, CK18 and α-SMA. ( E ) Serum supplementation and withdrawal of growth factors induced the expression of lineage specific markers of the mature mammary epithelium (CK14, CK18 and α-SMA) in cultured cells from mammosphere clones, with loss of the stem cell markers Oct-4 and nestin. ( F ) Representative FACS analysis of epithelial differentiated cells showing the acquirement of CD24, the expression of pan-CK and CK18 and the absence of the endothelial cell markers KDR and CD31. The dark area indicates the specific Ab, the white area the isotypic control. Original magnification: panel A: ×100; panel D and E: ×650. Nuclei were counterstained with Hoechst dye.

Article Snippet: The following antibodies were used: rabbit anti-von Willebrand Factor (vWF) Ab and anti-α-smooth muscle actin (α-SMA) mAb (Dako), rabbit anti-pan-CK Ab, goat anti-CK14, rabbit anti-CK18 Abs (Santa Cruz), anti-vimentin mAb (Sigma, clone 13.2), goat anti-Oct-4 Ab (Abcam, Cambridge, UK) and rabbit anti-nestin Ab (Chemicon, Temecula, CA, USA).

Techniques: Staining, Clone Assay, Generated, Expressing, Control, Immunofluorescence, Cell Culture

Journal: Journal of Cellular and Molecular Medicine

Article Title: Endothelial cell differentiation of human breast tumour stem/progenitor cells

doi: 10.1111/j.1582-4934.2008.00338.x

Figure Lengend Snippet: Endothelial differentiation of mammosphere clones cultured in endothelial differentiating medium containing VEGF. ( A ) Representative micrographs showing the expression of endothelial markers after 2-week culture by immunofluorescence staining of vascular endothelial (VE)-cadherin and vWF, and representative FACS analysis (KDR, CD105, CD31, CD146, VEGF receptor 3 (VEGFR3): black area; the white area is the isotypic control) showing the endothelial differentiation. CK18 was negative. ( B ) Representative micrographs showing the formation of capillary-like structures on Matrigel by endothelial-differentiated breast tumour stem cells, but not by undifferentiated breast tumour progenitor cells. Capillary-like formation assay was evaluated after 6 hrs. (Original magnification: panel A ×650; panel B ×200).

Article Snippet: The following antibodies were used: rabbit anti-von Willebrand Factor (vWF) Ab and anti-α-smooth muscle actin (α-SMA) mAb (Dako), rabbit anti-pan-CK Ab, goat anti-CK14, rabbit anti-CK18 Abs (Santa Cruz), anti-vimentin mAb (Sigma, clone 13.2), goat anti-Oct-4 Ab (Abcam, Cambridge, UK) and rabbit anti-nestin Ab (Chemicon, Temecula, CA, USA).

Techniques: Clone Assay, Cell Culture, Expressing, Immunofluorescence, Staining, Control, Tube Formation Assay

Journal: Stem Cell Research & Therapy

Article Title: Human umbilical cord-derived mesenchymal stem cells improve the function of liver in rats with acute-on-chronic liver failure via downregulating Notch and Stat1/Stat3 signaling

doi: 10.1186/s13287-021-02468-6

Figure Lengend Snippet: The effects of hUC-MSC transplantation on hepatocyte regeneration in ACLF rats. Liver sections from ACLF rats 12 and 24 h post-hUC-MSC transplantation or 0.9% sodium chloride injection as a control were used for immunohistochemical staining of AFP, CK18, HGF, and PCNA and microscopic examination ( n = 4/group). Representative photographs are shown ( a ). Positive staining was quantified and is presented as the mean ± SD ( b, c ). d Serum levels of HGF were detected by ELISA (4h, n = 3/group; 12h and 24h, n = 4/group). Data are presented as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The following primary antibodies were used: rabbit anti-alpha smooth muscle actin (α-SMA) mAb (1:200, Abcam, UK, ab32575), rabbit anti-desmin pAb (1:200, Abcam, UK, ab15200), rabbit anti-matrix metalloproteinase 9 (MMP9) mAb (1:1000, Abcam, UK, ab76003), rabbit anti-CD90 mAb (1:50, Abcam, UK, ab133350), rabbit anti-tissue inhibitor of metalloproteinase 1 (TIMP1) pAb (1:100, Proteintech, China, 16644-1-AP), rabbit anti-cytokeratin 18 (CK18) pAb (1:200, Proteintech, China, 10830-1-AP), rabbit anti-alpha fetoprotein (AFP) pAb (1:100, Affinity, USA, AF5134), rabbit anti-HGF pAb (1:100, Affinity, USA, DF6326), rabbit anti-proliferating cell nuclear antigen (PCNA) pAb (1:100, Affinity, USA, AF0239), rabbit anti-Phospho-Stat1 mAb (1:100, Cell Signaling Technology, USA, 8826S), and rabbit anti-Phospho-Stat3 mAb (1:100, Cell Signaling Technology, USA, 9145S).

Techniques: Transplantation Assay, Injection, Control, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Ex Vivo Stromal Cell-Derived Factor 1-Mediated Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells into Hepatocytes Is Enhanced by Chinese Medicine Yiguanjian Drug-Containing Serum

doi: 10.1155/2016/7380439

Figure Lengend Snippet: ALB and CK-18 protein expressions in differentiated BM-derived MSCs (P2). MSCs were cultured in the presence of 20 ng/mL HGF + 15% normal serum (a), 20 ng/mL HGF + 50 ng/mL SDF-1 + 15% normal serum (b), 20 ng/mL HGF + 50 ng/mL SDF-1 + 20% Yiguanjian drug-containing serum (c), or 20% Yiguanjian drug-containing serum (d), and protein expression was determined by western blot. β -actin was used as an inner control.

Article Snippet: For immunocytochemistry, cells were incubated with the following antibodies: rabbit anti-mouse ALB (1 : 200), rabbit anti-mouse CK-18 (1 : 200) (Bioss Biotechnology Co., Ltd., Beijing, China) overnight at 4°C followed by DAB staining.

Techniques: Derivative Assay, Cell Culture, Expressing, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Ex Vivo Stromal Cell-Derived Factor 1-Mediated Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells into Hepatocytes Is Enhanced by Chinese Medicine Yiguanjian Drug-Containing Serum

doi: 10.1155/2016/7380439

Figure Lengend Snippet: Cytokeratin-18 (CK-18) expression in differentiated BM-derived MSCs (P2). BM-MSCs were cultured in the presence of 20 ng/mL HGF + 15% normal serum (a), 20 ng/mL HGF + 50 ng/mL SDF-1 + 15% normal serum (b), 20 ng/mL HGF + 50 ng/mL SDF-1 + 20% YGJ (c), or 20% YGJ (d), and CK-18 was visualized by ICC at the indicated time points (magnification ×40: (a): (b2)-(b3) and (c): (b1)-(b2); magnification ×20: (a): (b1) and (b4), (b, c): (b3)-(b4), and (d)). CK-18 staining was observed at day 14 of culture in HGF + normal serum-treated BM-MSCs, and CK-18 staining density increased until day 28 (a). CK-18 staining was observed at day 7 of culture in HGF + SDF-1 + normal serum-treated BM-MSCs, and CK-18 staining density increased until day 28 (b). CK-18 staining was observed at day 7 of culture in HGF + SDF-1 + YGJ-treated BM-MSCs, and CK-18 staining density increased until day 28 (c). Positive staining for CK-18 was observed at day 14 in cells cultured with YGJ, and staining intensity increased until day 28 (d).

Article Snippet: For immunocytochemistry, cells were incubated with the following antibodies: rabbit anti-mouse ALB (1 : 200), rabbit anti-mouse CK-18 (1 : 200) (Bioss Biotechnology Co., Ltd., Beijing, China) overnight at 4°C followed by DAB staining.

Techniques: Expressing, Derivative Assay, Cell Culture, Staining