A novel method of neural differentiation of PC12 cells by using Opti-MEM as a basic induction medium

Neuritogenesis is the process underling nervous system regeneration; however, optimal extracellular signals that can promote neuronal regenerative activities require further investigation. Previously, we developed a novel method for inducing neuronal differentiation in rat PC12 cells using temperature-controlled repeated thermal stimulation (TRTS) with a heating plate. Based on neurogenic sensitivity to TRTS, PC12 cells were classified as either hyper- or hyposensitive. In this study, we aimed to investigate the mechanism of hyposensitivity by establishing two PC12-derived subclones according to TRTS sensitivity during differentiation: PC12-P1F1, a hypersensitive subclone, and PC12-P1D10, a hyposensitive subclone. To characterize these subclones, cell size and neuritogenesis were evaluated in subclones treated with nerve growth factor (NGF), bone morphogenetic protein (BMP), or various TRTS. No significant differences in cell size were observed among the parental cells and subclones. BMP4- or TRTS-induced neuritogenesis was increased in PC12-P1F1 cells compared to that in the parental cells, while no neuritogenesis was observed in PC12-P1D10 cells. In contrast, NGF-induced neuritogenesis was observed in all three cell lines. Furthermore, a BMP inhibitor, LDN-193189, considerably inhibited TRTS-induced neuritogenesis. These results suggest that the BMP pathway might be required for TRTS-induced neuritogenesis, demonstrating the useful aspects of these novel subclones for TRTS research.

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Lead Exposure in Pheochromocytoma (PC12) Cells Alters Neural Differentiation and Sp1 DNA-Binding

NeuroToxicology ◽

10.1016/s0161-813x(00)00008-5 ◽

2001 ◽

Vol 22 (1) ◽

pp. 49-62 ◽

Cited By ~ 42

Author(s):

T. Crumpton ◽

D.S. Atkins ◽

N.H. Zawia ◽

S. Barone

Keyword(s):

Dna Binding ◽

Pc12 Cells ◽

Lead Exposure ◽

Neural Differentiation ◽

Pheochromocytoma Pc12 ◽

Pheochromocytoma Pc12 Cells

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Evaluation of anti-Fas ligand-induced apoptosis and neural differentiation of PC12 cells treated with nerve growth factor using small interfering RNA method and sampling by microdialysis

Analytical Biochemistry ◽

10.1016/j.ab.2007.01.012 ◽

2007 ◽

Vol 363 (1) ◽

pp. 46-57 ◽

Cited By ~ 11

Author(s):

Shih-Hwa Chiou ◽

Chung-Lan Kao ◽

Yu-Lih Chang ◽

Hung-Hai Ku ◽

Yung-Jen Tsai ◽

...

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Pc12 Cells ◽

Small Interfering Rna ◽

Neural Differentiation ◽

Fas Ligand ◽

Nerve Growth ◽

Induced Apoptosis ◽

Interfering Rna

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Vimentin is important in the neural differentiation of PC12 cells promoted by sialylation

Glycoconjugate Journal ◽

10.1007/s10719-016-9727-6 ◽

2016 ◽

Vol 34 (1) ◽

pp. 51-59 ◽

Cited By ~ 3

Author(s):

Li Hui Wu ◽

Xiao Ting Shao ◽

Jun Xia Guo ◽

Hui Sun ◽

Qi Chen ◽

...

Keyword(s):

Pc12 Cells ◽

Neural Differentiation

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Human Adipose Derived Stem Cell Exosomes Enhance the Neural Differentiation of PC12 Cells

10.21203/rs.3.rs-191113/v1 ◽

2021 ◽

Author(s):

Samira Shariati Najafabadi ◽

Noushin Amirpour ◽

Sharhram Amini ◽

Nasrin Zare ◽

Mohammad Kazemi ◽

...

Keyword(s):

Cell Viability ◽

Acridine Orange ◽

Pc12 Cells ◽

Mtt Assay ◽

Neural Differentiation ◽

Treated Group ◽

Target Cells ◽

Therapeutic Effects ◽

Acridine Orange Staining ◽

Qrt Pcr

Abstract Background: Human adipose stem cells (hADSCs) are proper cell sources for tissue regeneration. They mainly mediate their therapeutic effects through paracrine factors as exosomes. The exosomes contents are protein, lipid and RNA. Exosomes are effective in restoring the function of neurons and astrocytes in neurodegenerative diseases, and improve the therapeutic outcomes. We investigated the effect of hADSCs derived exosomes on survival and neural differentiation of PC12 cells in vitro.Methods and Results: The isolated hADSCs, were characterized by flow cytometry. Exosomes were separated from hADSC-condition medium using Exo-spinTM kit and characterized by DLS and TEM. Then acridine orange staining was performed to confirm entrance of exosomes into PC12 cells. PC12 cells were treated with culture medium containing NGF and exosome. Cell viability was assessed by MTT assay, and neural differentiation by ICC technique and qRT-PCR. TEM and DLS data confirmed the isolation of exosomes according to their size (30-100nm) and acridine orange staining indicated entrance of exosomes to target cells. MTT assay showed that cell viability was significantly increased in exosome treated group. ICC technique revealed that the expression of Map2 was superior in the exosome treated group. Based on qRT-PCR data, Map2 and β-tub III gene expression was increased in the exosome treated group. Significant expression of Gfap was seen in the NGF and NGF/EXO treated groups.Conclusions: Present study indicated that hADSCs derived exosomes might enhance cell viability and promote neuronal differentiation and expression of mature neural marker in PC12 cells.

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Effects of Self-Assembling Peptide RADA16-І Hydrogel on Neural Differentiation of Bone Marrow Mesenchymal Stem Cells

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.998-999.238 ◽

2014 ◽

Vol 998-999 ◽

pp. 238-242 ◽

Cited By ~ 1

Author(s):

Yan Sheng Liao ◽

Li Deng ◽

Xiao Qing Gao ◽

Chao Xian Yang

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Neural Differentiation ◽

Fluorescent Protein ◽

Morphological Changes ◽

Control Group ◽

Induction Medium ◽

Self Assembling ◽

Marrow Mesenchymal Stem Cells

Objective: To establish composite culture system of rat bone marrow mesenchymal stem cells (BMSCs) and self-assembling peptide hydrogel RADA16-І, and to investigate the effect of RADA16-І hydrogel on neural differentiation of BMSCs. Methods: BMSCs were isolated, cultivated and labeled with green fluorescent protein (GFP), then they were inoculated on glass coverslips or in RADA16-І solution to form control group and RADA16-І group respectively. The morphological changes of BMSCs induced by neural induction medium were observed, and GFAP, NeuN and Map-2 expressions of BMSCs in each group were detected with immunofluorescence. Results: The induced BMSCs presented neuron-like change, and the rates of GFAP and NF-200 positive cells in RADA16-І group were higher than that in control group (P < 0.05). Conclusion: Self-assembling peptide RADA16-І hydrogel can promote neural differentiation of BMSCs, and which may be used as scaffold material on BMSCs transplantation for treatment of nervous system diseases.

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Breeding Rice lines for physio-functional food through indica ‘Zhaxima’ × japonica ‘Nanjing 46’ haploid technique

Food Production, Processing and Nutrition ◽

10.1186/s43014-019-0010-7 ◽

2019 ◽

Vol 1 (1) ◽

Author(s):

Q. Q. Tian ◽

X. Li ◽

C. M. Lu ◽

X. W. Fang

Keyword(s):

Oryza Sativa ◽

Rice Variety ◽

Short Chain Fatty Acids ◽

Basic Medium ◽

Eating Quality ◽

Induction Medium ◽

Efficient Technology ◽

Plant Materials ◽

Regenerated Plants ◽

Basic Induction

AbstractResistant starch (RS) encompasses those forms of starch which are not accessible to human digestive enzymes and are fermented in the colons producing short chain fatty acids. The plant materials containing RS are few in the world. In this contribution, the culture ability of callus from anthers of F1 plants from, landraces, ‘Zhaxima’(Oryza sativa var. indica, high-RS rice line with 7.705 ± 0.142, g/100 g) × ‘Nanjing 46’ (Oryza sativa var. japonica, rice variety with RS content (g/100 g) of 0.200 ± 0.001 crosses were studied for obtaining high RS rice plants. The results showed that when M8 basic induction medium was added with 1.5 mg /L 2,4-D、2 mg /LNAA and 0.3 mg /L KT, the inductivity of callus was high as 32.14% for 21 d after pretreatment at 4 °C for 3 d; When MS differentiation basic medium was added with 2 mg /LKT and 3 mg /L ABA, the frequency of regeneration for callus was 50.3% with only a regeneration frequency of 4.55% grown into green seedlings. The RS content in the seeds was between those of the two parents and was partially normally distributed, the highest RS contents of the regenerated plants was as high as 7.66 ± 1.197%. This produced an efficient technology for regenerating stable rice lines with high RS and good eating quality using anthers culture.

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