Neurogenic Differentiation of hPDLSCs

CTRL and MORINGIN-hPDLSCs were cultured in undifferentiated and neurogenic differentiated medium for 10 days. Differentiation medium was composed by Neurobasal-A Medium(Gibco®) containing B27 (2%), L-glutamine (2 mM), penicillin (100 U/ml), streptomycin (100 mg/ml) and amphotericin B (5 mg/ml) and supplemented with basic Fibroblast Growth Factor (bFGF, 20 ng/ml) (TemaRicerca, Milan, Italy)^{39 (link)}. Fixed cells were incubated with rabbit primary monoclonal antibody, anti-GAP-43 (rabbit, 1:200; Sigma Aldrich, Milan, Italy), anti-p75 (rabbit, 1:200; DBA, Milan, Italy), anti-BDNF (rabbit; 1:50; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-Nestin (rabbit, 1:200; Santa Cruz Biotechnology)^{40 (link),41 (link)}. Cells were incubated with anti-rabbit Alexa Fluor 568 (Molecular Probes, Life Technologies, Monza, MI, Italy). All samples were incubated with Alexa Fluor 488 phalloidin green fluorescence conjugate (1:400), to mark the cytoskeleton actin and with TOPRO to staining nuclei. Confocal laser scanning microscopy (Zeiss LSM800, Zeiss, Jena, Germany), equipped with a Plan Neofluar oil-immersion objective (40×/1.3 NA) was used to samples observation.

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Romeo L., Diomede F., Gugliandolo A., Scionti D., Lo Giudice F., Lanza Cariccio V., Iori R., Bramanti P., Trubiani O, & Mazzon E. (2018). Moringin Induces Neural Differentiation in the Stem Cell of the Human Periodontal Ligament. Scientific Reports, 8, 9153.

Publication 2018

Neurobasal-A Medium anti-GAP-43 anti-BDNF anti-Nestin Alexa Fluor 568 Zeiss LSM800 Plan Neofluar oil-immersion objective

Actin Alexa 568 Alexa fluor 488 Amphotericin b Basic fibroblast growth factor Cells Confocal laser scanning microscopy Cytoskeleton Fluorescence Gap 43 Immersion L glutamine Molecular probes Monoclonal antibody Moringin Nestin Neurogenic Nuclei Penicillin Phalloidin Rabbit Streptomycin

Corresponding Organization :

Other organizations : Centro Neurolesi Bonino Pulejo, University of Chieti-Pescara, Centro Agricoltura Ambiente (Italy)

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Variable analysis

independent variables

CTRL and MORINGIN-hPDLSCs cell types

dependent variables

Expression of GAP-43, p75, BDNF, and Nestin

control variables

Undifferentiated and neurogenic differentiated medium
Neurobasal-A Medium with B27, L-glutamine, penicillin, streptomycin, and amphotericin B
Basic Fibroblast Growth Factor (bFGF, 20 ng/ml)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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