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