Comparison of Ectopic Gene Expression Methods in Rat Neural Stem Cells

Farhanullah²,

et al. 2015

ACS Chem. Neurosci.

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Identification of small molecules that direct neural stem cells (NSCs) into specific cell types would be helpful to understand the molecular mechanisms involved in regulation of NSC fate, and facilitate the development of therapeutic applications. In the current study, we developed and screened small molecules that can modulate the fate of NSCs that are derived from rat fetal cortex. Among these compounds, compounds 5 and 6 successfully differentiated NSCs into astrocytes and neurons, respectively. Compound 5 induced astrocytogenesis by increasing expression of interleukin-6, bone morphogenetic protein 2 and leukemia inhibitory factor and through consequent phosphorylation of signal transducer and activator of transcription 3 and Sma- and Mad-related protein 1/5/8 in NSCs. In addition, compound 5 increased the expression of fibroblast growth factor (FGF) 2 and FGF8 which may regulate the branching and morphology of astrocytes. Taken together, our results suggest that these small molecules can serve as a useful tool to study cell fate determination in NSCs and be used as an inexpensive alternative to cytokines to study mechanisms of astrocytogenesis.

“…The procedures for preparation of rat NSCs were described previously . Animals were treated according to Chung-Ang University and NIH standards of animal care.…”

Section: Methodsmentioning

confidence: 99%

“…The procedures for preparation of rat NSCs were described previously. 58 Animals were treated according to Chung-Ang University and NIH standards of animal care. Briefly, NSCs were isolated from the cortex of E14 Sprague−Dawley rat (Orient Bio Inc., Gyeonggi-do, Korea) embryos.…”

Section: ■ Methodsmentioning

confidence: 99%

Discovery of a Small Molecule that Enhances Astrocytogenesis by Activation of STAT3, SMAD1/5/8, and ERK1/2 via Induction of Cytokines in Neural Stem Cells

Farhanullah²,

et al. 2015

ACS Chem. Neurosci.

Self Cite

“…NSCs from the cortex of Sprague–Dawley rat (Orient Bio Inc., Gyeonggi-do, Korea) embryos at embryonic day 14 (E14) were isolated and cultured as previously described [ 36 ]. Animal experiments were performed in accordance with Chung-Ang University and NIH standards of animal care and approved by Chung-Ang University animal care and use committee (Permit Number: 13–0049, 2014–00032).…”

Section: Methodsmentioning

confidence: 99%

Kuwanon V Inhibits Proliferation, Promotes Cell Survival and Increases Neurogenesis of Neural Stem Cells

et al. 2015

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Neural stem cells (NSCs) have the ability to proliferate and differentiate into neurons and glia. Regulation of NSC fate by small molecules is important for the generation of a certain type of cell. The identification of small molecules that can induce new neurons from NSCs could facilitate regenerative medicine and drug development for neurodegenerative diseases. In this study, we screened natural compounds to identify molecules that are effective on NSC cell fate determination. We found that Kuwanon V (KWV), which was isolated from the mulberry tree (Morus bombycis) root, increased neurogenesis in rat NSCs. In addition, during NSC differentiation, KWV increased cell survival and inhibited cell proliferation as shown by 5-bromo-2-deoxyuridine pulse experiments, Ki67 immunostaining and neurosphere forming assays. Interestingly, KWV enhanced neuronal differentiation and decreased NSC proliferation even in the presence of mitogens such as epidermal growth factor and fibroblast growth factor 2. KWV treatment of NSCs reduced the phosphorylation of extracellular signal-regulated kinase 1/2, increased mRNA expression levels of the cyclin-dependent kinase inhibitor p21, down-regulated Notch/Hairy expression levels and up-regulated microRNA miR-9, miR-29a and miR-181a. Taken together, our data suggest that KWV modulates NSC fate to induce neurogenesis, and it may be considered as a new drug candidate that can regenerate or protect neurons in neurodegenerative diseases.

Biomolecules & Therapeutics

“…NSCs were cultured as previously described ( Kim et al ., 2013 ). Briefly, NSCs isolated from the cortex of E14 Sprague-Dawley rat embryos were dissociated, seeded onto tissue culture flasks (200,000 cells/ml), and expanded as neurospheres for 7 days in Dulbecco’s modified Eagle’s medium/F12 supplemented with 1% (v/v) PSA, 2% (v/v) B27 (all from Gibco, Grand Island, NY, USA), 20 ng/ml EGF and 20 ng/ml FGF2 (both from Chemicon, Temecula, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

An Aminopropyl Carbazole Derivative Induces Neurogenesis by Increasing Final Cell Division in Neural Stem Cells

Shin

Kong

Yoon

et al. 2015

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P7C3 and its derivatives, 1-(3,6-dibromo-9H-carbazol-9-yl)-3-(p-tolylamino)propan-2-ol (1) and N-(3-(3,6-dibromo-9H-carbazol-9-yl)-2-hydroxypropyl)-N-(3-methoxyphenyl)-4-methylbenzenesulfonamide (2), were previously reported to increase neurogenesis in rat neural stem cells (NSCs). Although P7C3 is known to increase neurogenesis by protecting newborn neurons, it is not known whether its derivatives also have protective effects to increase neurogenesis. In the current study, we examined how 1 induces neurogenesis. The treatment of 1 in NSCs increased numbers of cells in the absence of epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2), while not affecting those in the presence of growth factors. Compound 1 did not induce astrocytogenesis during NSC differentiation. 5-Bromo-2′-deoxyuridine (BrdU) pulsing experiments showed that 1 significantly enhanced BrdU-positive neurons. Taken together, our data suggest that 1 promotes neurogenesis by the induction of final cell division during NSC differentiation.