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:

Article Title: Studies of delayed systemic effects of ultraviolet B radiation (UVR) on the induction of contact hypersensitivity, 2. Evidence that interleukin-10 from UVR-treated epidermis is the critical mediator

doi: 10.1046/j.1365-2567.2000.00934.x

Figure Lengend Snippet: CH‐inducing capacity of dendritic cells obtained from lymph nodes from IL‐10‐treated mice. Panels of mice received IL‐10 (200 ng) injections intradermally into abdominal skin. Eight hours later, lymph nodes not draining the injected skin (all except axillary and inguinal) were removed. Lymphoid cell suspensions were separated on density gradients using Percoll solution 1·075 g/dl, layered over 1·035 g/dl solution and centrifuged. Interface cells were collected and placed in Petri dishes at 37° for 60 min, after which non‐adherent cells were discarded. The adherent cells were then incubated at 37° overnight and non‐adherent cells were collected as cell suspension enriched for DCs. As positive control, cells were prepared from lymph nodes of untreated mice. DC‐enriched suspensions derivatized with DNFB in vitro were injected into footpads of naive syngeneic mice (2 × 104 cells in 100 µl per mouse). Five days later CH was assayed as described in the legend to Fig. 1. Bars represent mean ear‐swelling responses ± SEM for groups of five mice each. Asterisks indicate mean responses significantly less than the positive control (P < 0·0005).

Article Snippet: The cells were then separated on density fractions using low‐density Percoll solution (Pharmacia & Upjohn, Peapack, WN) (1·035 g/dl), layered over a high‐density solution (1·075 g/dl), and centrifuged at 570 g for 20 min. Interface cells were collected and placed in 10‐ml Petri dishes at 37° for 60 min, after which non‐adherent cells were discarded.

Techniques: Injection, Incubation, Positive Control, In Vitro