Journal: PLoS ONE

Article Title: The Neuro-Glial Properties of Adipose-Derived Adult Stromal (ADAS) Cells Are Not Regulated by Notch 1 and Are Not Derived from Neural Crest Lineage

doi: 10.1371/journal.pone.0001453

Figure Lengend Snippet: Expression of neural stem cell markers in mouse ADAS grown in Mesencult and exposed to Neural Induction Medium (NIM).

Article Snippet: Following a second centrifugation step (1000×g for 4 minutes), cells were filtered through a Nytex filter and plated in 2 mL of one of the following growth media into a 35 mm 2 cell culture dish: mouse ADAS cells were grown in Medium A (NeuroCult Proliferation Medium), Medium B ([DMEM/F12, 100 U penicillin, 100 μg streptomycin, 2 mM L-glutamine, 1× N2 Supplement, EGF (10 ng/mL; R&D), FGF-2 (20 ng/mL; R&D), and mLIF (10 ng/mL; R&D)], or Medium C ([MesenCult Basal Media (Stem Cell Technologies) supplemented with MesenCult Proliferation Supplement (1∶5), 100 U penicillin and 100 μg streptomycin).

Techniques: Expressing

Journal: PLoS ONE

Article Title: The Neuro-Glial Properties of Adipose-Derived Adult Stromal (ADAS) Cells Are Not Regulated by Notch 1 and Are Not Derived from Neural Crest Lineage

doi: 10.1371/journal.pone.0001453

Figure Lengend Snippet: Expression of neural markers in MesenCult proliferation media (A, B) and after 4-hr (C, D) or 24-hr (E, F) exposure to Neural Induction Medium (NIM). NSE, Neuron Specific Enolase; GFAP, glial-fibrillary acidic protein; Tuj1, β-tubulin III. Scale bar in A–F = 110 um.

Article Snippet: Following a second centrifugation step (1000×g for 4 minutes), cells were filtered through a Nytex filter and plated in 2 mL of one of the following growth media into a 35 mm 2 cell culture dish: mouse ADAS cells were grown in Medium A (NeuroCult Proliferation Medium), Medium B ([DMEM/F12, 100 U penicillin, 100 μg streptomycin, 2 mM L-glutamine, 1× N2 Supplement, EGF (10 ng/mL; R&D), FGF-2 (20 ng/mL; R&D), and mLIF (10 ng/mL; R&D)], or Medium C ([MesenCult Basal Media (Stem Cell Technologies) supplemented with MesenCult Proliferation Supplement (1∶5), 100 U penicillin and 100 μg streptomycin).

Techniques: Expressing

Journal: PLoS ONE

Article Title: The Neuro-Glial Properties of Adipose-Derived Adult Stromal (ADAS) Cells Are Not Regulated by Notch 1 and Are Not Derived from Neural Crest Lineage

doi: 10.1371/journal.pone.0001453

Figure Lengend Snippet: Nestin expression in floxed Notch 1 ADAS cells in mock-transduced (A, B), transduced with lentiviral-Cre (C, D), or lentiviral-mutant Cre (E, F) in MesenCult (A, C, E) was decreased by 70% after exposure to NeuroCult Differentiation Medium (B, D, F). as measured by optical density analyses (See Methods). Scale bar in A–F = 55 um.

Article Snippet: Following a second centrifugation step (1000×g for 4 minutes), cells were filtered through a Nytex filter and plated in 2 mL of one of the following growth media into a 35 mm 2 cell culture dish: mouse ADAS cells were grown in Medium A (NeuroCult Proliferation Medium), Medium B ([DMEM/F12, 100 U penicillin, 100 μg streptomycin, 2 mM L-glutamine, 1× N2 Supplement, EGF (10 ng/mL; R&D), FGF-2 (20 ng/mL; R&D), and mLIF (10 ng/mL; R&D)], or Medium C ([MesenCult Basal Media (Stem Cell Technologies) supplemented with MesenCult Proliferation Supplement (1∶5), 100 U penicillin and 100 μg streptomycin).

Techniques: Expressing, Transduction, Mutagenesis

Journal: PLoS ONE

Article Title: The Neuro-Glial Properties of Adipose-Derived Adult Stromal (ADAS) Cells Are Not Regulated by Notch 1 and Are Not Derived from Neural Crest Lineage

doi: 10.1371/journal.pone.0001453

Figure Lengend Snippet: Adipose from Wnt1-Cre;Rosa26R-stop-LacZ compound transgenic mice was harvested to establish ADAS cell cultures. X-gal enzymatic activity revealed no detectable blue cells in ADAS cultures grown in NeuroCult (A); a few isolated cells harboring the lacZ transgene in ADAS cultures grown in Mesencult (B); lacZ-positive neural crest-derived cells in the bulb of a whisker hair follicle of a Wnt1-Cre;Rosa26RlacZ transgenic mouse prior to dissociation (C); lacZ-positive cells migrating out of the bulb sheath at the base of a whisker hair follicle in culture after dissociation (D); Phase image of ADAS cultures grown in Mesencult derived from Wnt1-Cre;Rosa26R-stop-lacZ compound transgenic mice (E); Immunofluorescence analysis with an antibody specific for bacterial β-galactosidase revealed the presence of a minimal number of double-labeled EYFP (yellow)/β-galactosidase (red) cells in these cultures (F), n = 3; Quantitative FACS analyses of ADAS cells derived from three different (G, H, I) Wnt1-Cre;Rosa26R-stop-EYFP compound transgenic mice revealed that less than 2% of the cells were EYFP-positive (F), n = 3. Scale bar in A–E = 55 um.

Article Snippet: Following a second centrifugation step (1000×g for 4 minutes), cells were filtered through a Nytex filter and plated in 2 mL of one of the following growth media into a 35 mm 2 cell culture dish: mouse ADAS cells were grown in Medium A (NeuroCult Proliferation Medium), Medium B ([DMEM/F12, 100 U penicillin, 100 μg streptomycin, 2 mM L-glutamine, 1× N2 Supplement, EGF (10 ng/mL; R&D), FGF-2 (20 ng/mL; R&D), and mLIF (10 ng/mL; R&D)], or Medium C ([MesenCult Basal Media (Stem Cell Technologies) supplemented with MesenCult Proliferation Supplement (1∶5), 100 U penicillin and 100 μg streptomycin).

Techniques: Transgenic Assay, Activity Assay, Isolation, Derivative Assay, Whisker Assay, Immunofluorescence, Labeling

Journal: The Journal of Experimental Medicine

Article Title: GM-CSF regulates intimal cell proliferation in nascent atherosclerotic lesions

doi: 10.1084/jem.20090866

Figure Lengend Snippet: PTx blockade of monocyte recruitment reveals that proliferating intimal cells accumulate in 2-wk lesions, and intimal cell proliferation is independent of monocyte recruitment. (a) Flow cytometry plots of peritoneal exudate cells harvested 24 h after i.p. thioglycollate injection of control and PTx-treated mice (percentages are indicated). Representative data from five mice per group studied in two independent experiments are shown. Additional details are provided in Fig. S2 . (b) Ldlr −/− mice fed a CRD for 2 wk were injected with BrdU at 2 h before treatment with PTx or PBS (C, control). Aortas were harvested 24 h (left) and 2 h (right) after BrdU injection, and BrdU + nuclei were enumerated. (c) Ldlr −/− mice fed a CRD for 2 wk were injected with PTx, and BrdU pulse labeling was performed 22 or 46 h later. Aortas were harvested after 2 h, and BrdU + nuclei were enumerated. In b and c, four independent experiments were performed at each time point, and means ± SEM were derived from four mice per group. *, P < 0.05 relative to C; **, P < 0.01.

Article Snippet: In BrdU labeling experiments, mice received a single 0.2-ml i.v. injection of 2 mg BrdU in PBS (BD).

Techniques: Flow Cytometry, Injection, Labeling, Derivative Assay

Journal: PLoS ONE

Article Title: Potential Relevance of α 1 -Adrenergic Receptor Autoantibodies in Refractory Hypertension

doi: 10.1371/journal.pone.0003742

Figure Lengend Snippet: (A) α 1 -AAB isolated from the serum of a patient with refractory hypertension elicited a Ca 2+ signal. (B) The rabbit α 1 -AB gave a similar Ca 2+ signal. (C) A human control IgG preparation was unable to affect intracellular Ca 2+ . Cardiomyocytes were electrically stimulated at 1 Hz, and the peak Ca 2+ was monitored.

Article Snippet: Following, vessel rings were incubated with α 1 -AAB (5 µg in 5 ml PBS buffer), rabbit α 1 -AB (50 µg in 5 ml PBS buffer), rabbit IgG (50 µg in 5 ml PBS buffer, Dunn Labortechnik GmBH, Asbach, Germany) and human control IgG endobulin (50 µg in 5 ml PBS buffer).

Techniques: Isolation

Journal: PLoS ONE

Article Title: Potential Relevance of α 1 -Adrenergic Receptor Autoantibodies in Refractory Hypertension

doi: 10.1371/journal.pone.0003742

Figure Lengend Snippet: A brisk KCl response is documented. (A) Representative experiment showing the contractile response to phenylephrine (PE, 10 nM–10 µM) and rabbit α 1 -AB (50 µg in 5 ml PBS buffer). The PE and antibody responses were similar in kind. (B) Contractile response of human α 1 -AAB isolated from two patients (5 µg in 5 ml PBS buffer) and rabbit α 1 -AB (50 µg in 5 ml PBS buffer) are compared to the PE response (300 nM and 10 µM, respectively) is shown. Contractions are expressed as % KCl response from patient 1 and patient 2. The human antibody responses were less than the low and high-dose PE responses. The rabbit α 1 -AB responses approached the PE responses. Two columns on the right are control human and control rabbit IgG showing no response.

Article Snippet: Following, vessel rings were incubated with α 1 -AAB (5 µg in 5 ml PBS buffer), rabbit α 1 -AB (50 µg in 5 ml PBS buffer), rabbit IgG (50 µg in 5 ml PBS buffer, Dunn Labortechnik GmBH, Asbach, Germany) and human control IgG endobulin (50 µg in 5 ml PBS buffer).

Techniques: Isolation