Name: modeling astrocytoma pathogenesis in vitro and in vivo using cortical astrocytes or neural stem cells from conditional, genetically engineered mice
Uploaded: 2014-08-12T15:00:00
Description: Watch this Scientific Journal Video about Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice at JoVE.co

CRM is a Damon Runyon-Genentech Clinical Investigator. This work was supported, in part, by grants to CRM from the Damon Runyon Cancer Research Foundation (CI-45-09), Department of Defense (W81XWH-09-2-0042), and University of North Carolina University Cancer Research Fund (UCRF). The authors wish to thank Daniel Roth for mouse husbandry assistance. The authors also wish to thank Hannah Chae, Carter McCormick, Demi Canoutas, Stephanie Gillette, and Susannah Krom for tissue culture and immunofluorescence assistance.

सभी जानवरों के अध्ययन उत्तरी केरोलिना संस्थागत पशु की देखभाल और उपयोग समिति के विश्वविद्यालय द्वारा अनुमोदित किया गया. नवजात चूहों से 1 संवर्धन Cortical Astrocytes <ol><li> तैयारी <ol><li> 2-3 नाजुक कार्य ऊतकों प?...

सबसे महत्वपूर्ण कदम, 2), 3) अच्छी तरह से कोशिकाओं को अलग कर देना, तानिका दूर करने के लिए महासंयोजिका नीचे ऊतक लेने के बिना प्रांतस्था आबकारी करने के लिए उचित फसल और cortical astrocytes की संस्कृति हैं 1) सुनिश्चित करने ?...

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			<td style="width:233px;">DMEM can also be purchased from other suppliers including GIBCOSigma and Cellgro</td>
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			<td style="width:233px;">Other Trypsin solutions are also suitable for cortical astrocyte havest and culture</td>
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modeling astrocytoma pathogenesis in vitro and in vivo using cortical astrocytes or neural stem cells from conditional, genetically engineered mice

Current astrocytoma models are limited in their ability to define the roles of oncogenic mutations in specific brain cell types during disease pathogenesis and their utility for preclinical drug development. In order to design a better model system for these applications, phenotypically wild-type cortical astrocytes and neural stem cells (NSC) from conditional, genetically engineered mice (GEM) that harbor various combinations of floxed oncogenic alleles were harvested and grown in culture. Genetic recombination was induced in vitro using adenoviral Cre-mediated recombination, resulting in expression of mutated oncogenes and deletion of tumor suppressor genes. The phenotypic consequences of these mutations were defined by measuring proliferation, transformation, and drug response in vitro. Orthotopic allograft models, whereby transformed cells are stereotactically injected into the brains of immune-competent, syngeneic littermates, were developed to define the role of oncogenic mutations and cell type on tumorigenesis in vivo. Unlike most established human glioblastoma cell line xenografts, injection of transformed GEM-derived cortical astrocytes into the brains of immune-competent littermates produced astrocytomas, including the most aggressive subtype, glioblastoma, that recapitulated the histopathological hallmarks of human astrocytomas, including diffuse invasion of normal brain parenchyma. Bioluminescence imaging of orthotopic allografts from transformed astrocytes engineered to express luciferase was utilized to monitor in vivo tumor growth over time. Thus, astrocytoma models using astrocytes and NSC harvested from GEM with conditional oncogenic alleles provide an integrated system to study the genetics and cell biology of astrocytoma pathogenesis in vitro and in vivo and may be useful in preclinical drug development for these devastating diseases.

हम phenotypically इन विट्रो में और vivo में लक्षण वर्णन किया जा सकता है कि सशर्त oncogenic alleles (चित्रा 1) floxed नवजात मणि को शरण देने से काटा कॉर्टिकल astrocytes और एनएससी के साथ एक nGEM मॉडल प्रणाली विकसित की है. विशेष रूप स?...

Watch this Scientific Journal Video about Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice at JoVE.co

Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice

मॉडलिंग तारिकाकोशिकार्बुद रोगजनन<em&gt; इन विट्रो</em&gt; और<em&gt; इन विवो</em&gt; Cortical Astrocytes या सशर्त, आनुवंशिक रूप से इंजीनियर चूहों से तंत्रिका स्टेम कोशिकाओं का उपयोग

Phenotypically wild-type astrocytes and neural stem cells harvested from mice engineered with floxed, conditional oncogenic alleles and transformed via viral Cre-mediated recombination can be used to model astrocytoma pathogenesis in vitro and in vivo by orthotopic injection of transformed cells into brains of syngeneic, immune-competent littermates.

Current astrocytoma models are limited in their ability to define the roles of oncogenic mutations in specific brain cell types during disease ...

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Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice

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Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

Expansion of Embryonic and Adult Neural Stem Cells by In Utero Electroporation or Viral Stereotaxic Injection

Expansion of Embryonic and Adult Neural Stem Cells by In Utero Electroporation or Viral Stereotaxic Injection

भ्रूण और वयस्क तंत्रिका स्टेम कोशिकाओं के द्वारा विस्तार<em&gt; Utero में</em&gt; Electroporation या वायरल Stereotaxic इंजेक्शन

Somatic stem cells can divide to generate additional stem cells (expansion) or more differentiated cell types (differentiation), which is fundamental for ...

Isolation and Culture of Neural Crest Cells from Embryonic Murine Neural Tube

अलगाव और भ्रूण murine तंत्रिका ट्यूब से तंत्रिका शिखा कोशिकाओं की संस्कृति

The embryonic neural crest (NC) is a multipotent progenitor population that originates at the dorsal aspect of the neural tube, undergoes an epithelial to ...

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

Vivo में दो photon इमेजिंग cortical न्यूरॉन्स में अनुभव पर निर्भर आणविक परिवर्तन

The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety ...

A cGMP-applicable Expansion Method for Aggregates of Human Neural Stem and Progenitor Cells Derived From Pluripotent Stem Cells or Fetal Brain Tissue

मानव तंत्रिका के समुच्चय के लिए एक cGMP-लागू विस्तार विधि स्टेम और पूर्वपुस्र्ष कोशिकाओं pluripotent स्टेम सेल या भ्रूण के मस्तिष्क के ऊतकों से निकाली

A cell expansion technique to amass large numbers of cells from a single specimen for research experiments and clinical trials would greatly benefit the ...

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators

इमेजिंग Intracellular सीए<sup&gt; 2 +</sup&gt; आनुवंशिक रूप से इनकोडिंग कैल्शियम संकेतकों का उपयोग वयस्क चूहों से Striatal astrocytes में सिग्नल

Astrocytes display spontaneous intracellular Ca2+ concentration fluctuations ([Ca2+]i) and in several settings respond to neuronal excitation with ...

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

पृष्ठीय रूट नाड़ीग्रन्थि आदर्श: कैंसर तंत्रिका आक्रमण की इन विट्रो मॉडलिंग में

One way that solid tumors disseminate is through neural invasion. This route is well-known in cancers of the head and neck, prostate, and pancreas. These ...

The Detection of 5-Hydroxymethylcytosine in Neural Stem Cells and Brains of Mice

तंत्रिका स्टेम सेल और चूहे के दिमाग में 5-hydroxymethylcytosine का पता लगाने

Multiple&#160;DNA modifications have been identified in the mammalian genome. Of that, 5-methylcytosine and 5-hydroxymethylcytosine-mediated epigenetic ...