adherent pc12 cells  (ATCC)

Journal: Scientific Reports

Article Title: Hyperglycemia-induced mitochondrial abnormalities in autonomic neurons via the RAGE axis

doi: 10.1038/s41598-025-10933-y

Figure Lengend Snippet: Detection of RAGE protein in mitochondria-enriched samples from PC12 NGF cells. A-B , Immunoblots and quantification of the level of expression of the neuronal markers MAP-2, neurofilament light chain (NF-L) and tubulin in whole PC12 samples maintained in media without (-NGF) or with NGF added (+ NGF). C , The immunoblots show the levels of RAGE, VDAC and tubulin detected in the cytosol-enriched and mitochondria-enriched samples from PC12 cells transformed by NGF (PC12 NGF ) maintained in either control (CTL) or high glucose (HG) conditions. C-D , The bar graphs show mean ± SEM levels of each protein after normalization to tubulin or VDAC in cytosol-enriched and mitochondria-enriched samples, respectively. Means were statistically compared by the Mann-Whitney U test; (MAP2: t 4 = 2.695; NF-L: t 4 = 3.297); * p < 0.05. ( N = 3 per group in A and C). For clarity, the blots shown in A and C have been cropped, and the background brightness has been adjusted (complete blots available in Supplementary Data file).

Article Snippet: Adherent PC12 cells (ATCC ® CRL-1721.1TM) were cultured in F-12 K growth media (2.5% fetal bovine serum, 15% horse serum; PC12 GM).

Techniques: Western Blot, Expressing, Transformation Assay, Control, MANN-WHITNEY

Journal: Foods

Article Title: Bioinformatics and Deep Learning Approach to Discover Food-Derived Active Ingredients for Alzheimer’s Disease Therapy

doi: 10.3390/foods14010127

Figure Lengend Snippet: Cell viability ( A ) and acetylcholinesterase (AChE) activity ( B ) Nerve growth factor (NGF)-differentiated PC12 cells with lipopolysaccharide-induced inflammation (LPS, 1 μg/mL) were treated with astragalin, dihydromyricetin, coumarin, quercetin, kaempferol, apigenin, and luteolin for 24 h to measure cell viability and 48 h to measure AChE activity. a–e Different letters on the bar indicated significant differences between the groups at p < 0.05.

Article Snippet: The cells of the PC12 adherent cell line (ATCC CRL-1721.1) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Sigma, Aldrich, St. Louis, MO, USA), supplemented with 10% horse serum, 5% fetal bovine serum, and 1% antibiotic mixture containing penicillin–streptomycin, in a humidified atmosphere at 37 °C with 5% CO 2 .

Techniques: Activity Assay

Journal: Foods

Article Title: Bioinformatics and Deep Learning Approach to Discover Food-Derived Active Ingredients for Alzheimer’s Disease Therapy

doi: 10.3390/foods14010127

Figure Lengend Snippet: Lipid peroxidation ( A ) and protein ( B ) and mRNA expression ( C ) of pro-inflammatory cytokines. Nerve growth factor (NGF)-differentiated PC12 cells with lipopolysaccharide-induced inflammation (LPS, 1 μg/mL) were treated with astragalin, dihydromyricetin, coumarin, quercetin, kaempferol, apigenin, and luteolin for 48 h. a–e Different letters on the bar indicated significant differences between the groups at p < 0.05.

Article Snippet: The cells of the PC12 adherent cell line (ATCC CRL-1721.1) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Sigma, Aldrich, St. Louis, MO, USA), supplemented with 10% horse serum, 5% fetal bovine serum, and 1% antibiotic mixture containing penicillin–streptomycin, in a humidified atmosphere at 37 °C with 5% CO 2 .

Techniques: Expressing

Journal: Scientific Reports

Article Title: Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents

doi: 10.1038/s41598-021-87431-4

Figure Lengend Snippet: Optimisation of media for 3 PC12 cell variants. Cells were adapted from their original media as described in “ ”. Representative phase contrast images are shown after 3 passages in the new media and imaged 2–3 days after passaging. All cells were supplemented with 15% horse serum and 2.5% foetal bovine serum. ( A ) PC12 Adh cells were adapted from Ham’s F-12K to DMEM and RPMI, with no differences seen in cell morphology. ( B ) PC12 cells Riken were adapted from DMEM to RPMI resulting in rounded, phase-bright cells with loss of cell-substratum adhesion. ( C ) NS-1 cells adapted from RPMI to DMEM showed increased neurite outgrowth. Scale bar ( A ) 200 µm. ( B , C ) 100 µm.

Article Snippet: Non-adherent PC12 cells (CRL-1721) from ATCC had been previously studied , , .

Techniques: Passaging

Journal: Scientific Reports

Article Title: Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents

doi: 10.1038/s41598-021-87431-4

Figure Lengend Snippet: Optimisation of substratum coating for 3 PC12 variants. PC12 cells were cultured in media with 15% horse serum and 2.5% foetal bovine serum ( A , C , E ) or serum-free supplemented media ( B , D ) as described in “ ”. Cells were passaged into 6-well plates with the wells either uncoated (non) or coated with collagen I (I), collagen IV (IV), poly- d -lysine (PDL), poly- l -lysine (PLL) or laminin (LM). Representative phase contrast photomicrographs were imaged after 24 h. ( A , B ) PC12 Adh cells cultured in Ham’s F-12 K medium were passaged into a 6-well plate with the indicated substratum coating with serum ( A ) or without serum ( B ). ( C , D ) PC12 Riken cells cultured in DMEM medium were passaged into wells with the indicated substratum coatings with serum ( C ) or without serum ( D ). ( E ) NS-1 cells cultured in DMEM medium with serum were passaged into a 6-well plate having substrata with the indicated coatings. Scale bar ( A , B ) 200 µm. ( C – E ) 100 µm.

Article Snippet: Non-adherent PC12 cells (CRL-1721) from ATCC had been previously studied , , .

Techniques: Cell Culture

Journal: Scientific Reports

Article Title: Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents

doi: 10.1038/s41598-021-87431-4

Figure Lengend Snippet: NGF differentiation of 3 PC12 cell variants. PC12 cells were cultured as described in “ ” and seeded into multiwell plates. After an overnight incubation, the media was changed to one with 2% horse serum (1% for PC12 Adh) and NGF added to the indicated concentration (ng/ml). The media and NGF was refreshed every 48 h and neurite scoring performed at 96 h. ( A ) PC12 Adh cells cultured in Ham’s F-12 K with serum were seeded into a poly- d -lysine-coated 6-well plate at 3 × 10 3 cells/cm 2 . Representative phase contrast images at 100 × magnification were taken 96 h after addition of the indicated concentration of NGF (ng/ml). ( B ) PC12 Riken cells cultured in DMEM with serum were plated onto a 12-well plate coated with collagen IV at a density of 4 × 10 3 cells/cm 2 . Representative phase contrast images at 200X magnification are shown 96 h after addition of NGF at the indicated concentrations (ng/ml). ( C ) NS-1 cells cultured in DMEM with serum were seeded into a collagen-IV coated 12-well plate at a density of 4 × 10 3 cells/cm 2 . Representative phase contrast images (200X magnification) are shown 96 h after addition of NGF at the indicated concentration (ng/ml). ( D ) Three PC12 variants were treated with NGF at the indicated concentrations and the percentage of neurite-bearing cells obtained after 96 h. Data shown is the mean and ± SEM of 3 independent experiments. Scale bar ( A ) 200 µm, ( B , C ) 100 µm.

Article Snippet: Non-adherent PC12 cells (CRL-1721) from ATCC had been previously studied , , .

Techniques: Cell Culture, Incubation, Concentration Assay

Journal: Scientific Reports

Article Title: Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents

doi: 10.1038/s41598-021-87431-4

Figure Lengend Snippet: Optimisation and validation of PC12 NS-1 in vitro stroke model. ( A ) NGF-differentiated NS-1 cells were subjected to OGD as described under “ ”. Caspase 3/7 activity was measured at the indicated durations of OGD and data normalised as fold increase over cells without OGD. Data shown is mean ± SEM from 3 independent experiments of triplicate determination. Statistical testing was performed with one-way ANOVA with Dunnett’s post hoc test. ***p < 0.001 compared to non-OGD control ( B ) NS-1 cells differentiated with NGF were subjected to OGD as described under “ ” for the indicated durations. Cell viability was then assessed using the MTT assay and data normalised to control cells without OGD. Data are mean ± SEM of 3 independent experiments carried out in triplicate. ***p < 0.001 compared to non-OGD control. Differentiated NS-1 cells were subjected to 3 h OGD then 24 h of normal culture conditions added with WAY100635 or 8-OH-DPAT (or both) as described under “ ”. Subsequently, cell viability or apoptosis was measured. ( C ) Cell viability was determined with MTT assay. *p < 0.05 compared to non-OGD control. ( D ) Caspase 3/7 activity was measured. Non-OGD baseline activity was removed and data normalised to untreated cells. **p < 0.01 compared to untreated control. All data are expressed as mean ± SEM of 3 experiments determined in triplicate.

Article Snippet: Non-adherent PC12 cells (CRL-1721) from ATCC had been previously studied , , .

Techniques: Biomarker Discovery, In Vitro, Activity Assay, Control, MTT Assay

Journal: Scientific Reports

Article Title: Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents

doi: 10.1038/s41598-021-87431-4

Figure Lengend Snippet: Application of PC12 NS-1 in vitro stroke model. ( A ) After 3-h OGD, NGF-differentiated NS-1 cells were incubated at normal conditions and treated with the indicated drugs as described under “ ”. Cell viability was determined by MTT assay. Data was normalised to untreated control cells. ( B ) Differentiated, non-OGD PC12 NS-1 cells were treated with the indicated drugs as described under “ ”. Cell viability was measured with MTT assay. Data was normalised to untreated control. ( C ) NGF-differentiated NS-1 cells were subjected to 3-h OGD followed by 24 h incubation at normal conditions with the indicated drugs as described under “ ”. Caspase 3/7 activity was measured. Non-OGD baseline activity was removed and data normalised to untreated cells. *p < 0.05 compared to control untreated cells; **p < 0.01 compared to control untreated cells. All data are expressed as mean ± SEM of 3 experiments determined in triplicate.

Article Snippet: Non-adherent PC12 cells (CRL-1721) from ATCC had been previously studied , , .

Techniques: In Vitro, Incubation, MTT Assay, Control, Activity Assay