Journal: Frontiers in Oncology

Article Title: Nestin(+) Tissue-Resident Multipotent Stem Cells Contribute to Tumor Progression by Differentiating into Pericytes and Smooth Muscle Cells Resulting in Blood Vessel Remodeling

doi: 10.3389/fonc.2014.00169

Figure Lengend Snippet: Tissue-resident Nest-GFP(+) cells and not BM-derived MSCs contribute to remodeling of tumor blood vessels . (A) B16F10 tumor cells were subcutaneously transplanted into the flank of Nest-GFP mice. Tumors were removed 21 days after tumor induction and subjected to immunofluorescence analysis. Vessels were stained for CD31 (endothelial cells), smooth muscle actin (SMA; pericytes/SMC), and GFP or Nestin, respectively. Control sections were stained with isotype controls. Nest-Control, Nest-GFP transgenic mice. Scale bar 15 μm. (B) Immunofluorescence analysis of B16F10 tumors grown subcutaneously in the flank of BM-reconstituted Nest-GFP mice. NestwtBM, Nest-GFP mice received bone-marrow (BM) from wt (C57Bl/6) mice after lethally total body irradiation (7 + 3Gy). (C) Experiment with reciprocal BM chimera as shown in (B) . wtNestBM, wt mice received bone-marrow (BM) from Nest-GFP mice after lethally total body irradiation. Scale bar 20 μm. (D) GFP-expression was quantified by counting GFP- and SMA-immunoreactive vascular structures (pericytes/SMC-stabilized vessels). (E) GFP-expression was further analyzed using Western blot analysis.

Article Snippet: Nestin antibody was from Acris Antibodies (Herford, Germany), alpha smooth muscle actin (SMA) and CD34 antibodies were from Santa Cruz (Santa Cruz, USA), CD31 antibody was from Dianova (Hamburg, Germany), and GFP antibody was from Thermo Scientific (Dreieich, Germany).

Techniques: Derivative Assay, Immunofluorescence, Staining, Control, Transgenic Assay, Irradiation, Expressing, Western Blot

Journal: Frontiers in Oncology

Article Title: Nestin(+) Tissue-Resident Multipotent Stem Cells Contribute to Tumor Progression by Differentiating into Pericytes and Smooth Muscle Cells Resulting in Blood Vessel Remodeling

doi: 10.3389/fonc.2014.00169

Figure Lengend Snippet: Tissue-resident Nest-GFP(+) pericytes associate with angiogenic vessels in LLC tumors grown in Nestin-GFP mice . (A) Immunofluorescence analysis of LLC tumors grown subcutaneously in the flank of wt BM-reconstituted Nest-GFP mice (NestwtBM). Tumors were isolated at day 21 and subjected for immunofluorescence analysis. Vessels were stained for CD31 or SMA and GFP. (B) Experiment with reciprocal BM chimera as shown in (A) . wtNestBM, wt mice having received BM from Nest-GFP mice after total body irradiation. To further confirm that tissue-resident MSCs and not BM-derived MSCs contribute to vascular remodeling processes one group of irradiated animals received NestBM for hematopoietic reconstitution that was supplemented with BM-derived EGFP-labeled MSC. Control sections were stained with isotype controls. Scale bar 20 μm. (C,D) GFP-expression was quantified by counting GFP and SMA-immunoreactive vessels and further analyzed using Western blot analysis.

Article Snippet: Nestin antibody was from Acris Antibodies (Herford, Germany), alpha smooth muscle actin (SMA) and CD34 antibodies were from Santa Cruz (Santa Cruz, USA), CD31 antibody was from Dianova (Hamburg, Germany), and GFP antibody was from Thermo Scientific (Dreieich, Germany).

Techniques: Immunofluorescence, Isolation, Staining, Irradiation, Derivative Assay, Labeling, Control, Expressing, Western Blot