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: Biology of sex differences

Article Title: Neuroprotective and neurotoxic outcomes of androgens and estrogens in an oxidative stress environment.

doi: 10.1186/s13293-020-0283-1

Figure Lengend Snippet: Fig. 1 Expression profile of hormone receptors. N27, PC12, C6, and MCF7 cells express the androgen receptor splice variant, AR45, and not full- length androgen receptors. In contrast, Jurkat cells express the full-length androgens receptor but not the AR45, which indicate that Jurkat cells are useful as both positive control and negative control for full-length androgen receptors and AR45, respectively (a, b). AR45 protein is expressed in human hippocampal tissue (c). Using the C terminus domain androgen receptor antibody, AR-C19, AR45 expression was detected in the membrane fraction and the whole cell lysate of frozen hippocampal postmortem tissue obtained from donors diagnosed with Alzheimer’s disease. No significant differences were observed between AR45 expression in whole cell lysate versus the membrane fraction, indicating AR45 is a membrane-associated androgen receptor. As expected, full-length androgen receptors (110 kDa) were not observed in frozen samples. However, an androgen receptor fragment (70 kDa) was observed in whole cell lysate, indicating the full-length androgen receptor that resides in the cytosol and nucleus was degraded to fragments. To confirm AR45 protein expression, which is missing the N terminus domain (NTD), the AR- N20 NTD targeted androgen receptor antibody was used. No AR45 expression was detected with AR-N20 antibody. Results were determined by ANOVA. Results are reported as mean + SEM. p < 0.05; WCL, whole cell lysate; MEM, membrane fraction; AR, androgen receptor

Article Snippet: 1RB3AN27 (N27) dopaminergic cells (kind gift from Randy Strong, Ph.D., at University of Texas Health Science Center; RRID: CVCL_D584), PC12 dopaminergic adherent cells (ATCC CRL-1721.1; RRID: CVCL_F659), and C6 glial cells (ATCC CCL-107; RRID: CVCL_0194) were used in experimental paradigms.

Techniques: Expressing, Variant Assay, Positive Control, Negative Control, Membrane

Journal: Biology of sex differences

Article Title: Neuroprotective and neurotoxic outcomes of androgens and estrogens in an oxidative stress environment.

doi: 10.1186/s13293-020-0283-1

Figure Lengend Snippet: Fig. 3 Effect of hormone pretreatment in PC12 cells in the presence of oxidative stress. Hormone alone has no effect on cell viability. H2O2 decreased cell viability by ~ 30%. Testosterone pretreatment prior to H2O2 is protective in PC12 cells. DHT-BSA, followed by H2O2 exposure, did not protect PC12 cells in an oxidative stress environment. Results were determined by ANOVA followed by Fisher LSD post hoc test. Results are reported as mean + SEM. p < 0.05; *versus control, **versus H2O2. C, vehicle control; T, 100 nM testosterone; H, H2O2; TH, pre-treatment T; D, 500 nM DHT-BSA; DH, pre-treatment D

Article Snippet: 1RB3AN27 (N27) dopaminergic cells (kind gift from Randy Strong, Ph.D., at University of Texas Health Science Center; RRID: CVCL_D584), PC12 dopaminergic adherent cells (ATCC CRL-1721.1; RRID: CVCL_F659), and C6 glial cells (ATCC CCL-107; RRID: CVCL_0194) were used in experimental paradigms.

Techniques: Control

Journal: Biology of sex differences

Article Title: Neuroprotective and neurotoxic outcomes of androgens and estrogens in an oxidative stress environment.

doi: 10.1186/s13293-020-0283-1

Figure Lengend Snippet: Fig. 7 Neurotoxic effects of hormones in an oxidative stress environment on PC12 cells. Alone, DHT-BSA had no effect on cell viability. H2O2 decreased cell viability by ~ 30%. DHT-BSA further decreased cell viability in the presence of oxidative stress in PC12 cells. Results were determined by ANOVA followed by Fisher LSD post hoc test. Results are reported as mean + SEM. p < 0.05; *versus control, **versus H2O2. C, vehicle control; H, H2O2; D, 500 nM DHT-BSA; HD, post-treatment D

Article Snippet: 1RB3AN27 (N27) dopaminergic cells (kind gift from Randy Strong, Ph.D., at University of Texas Health Science Center; RRID: CVCL_D584), PC12 dopaminergic adherent cells (ATCC CRL-1721.1; RRID: CVCL_F659), and C6 glial cells (ATCC CCL-107; RRID: CVCL_0194) were used in experimental paradigms.

Techniques: Control

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