Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

Eva Porlan; Beatriz Mart&#237;-Prado; Antonella Consiglio; Isabel Fari&#241;as

doi:10.3791/53282

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Nöral Analizi Yetişkin Fare V-SVZ'una Hücre istikrarlı ve verimli Genetik Modifikasyon Kök Hücre Özerk ve Non-özerk Etkileri

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Gelişim Biyolojisi

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JoVE Journal Gelişim Biyolojisi

Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

Nöral Analizi Yetişkin Fare V-SVZ'una Hücre istikrarlı ve verimli Genetik Modifikasyon Kök Hücre Özerk ve Non-özerk Etkileri

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9,157 Views

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08:48 min

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February 17, 2016

DOI:

10.3791/53282-v

DOI:

10.3791/53282-v

•

08:48 min

February 17, 2016

•

5 Views

Eva Porlan¹, Beatriz Martí-Prado^2,3, Antonella Consiglio^4,5, Isabel Fariñas^2,3

¹Cell Division and Cancer Group,Spanish National Cancer Research Centre (CNIO), ²Centro de Investigaciones Biomédicas en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ³Departmento de Biologìa Celular,Universidad de Valencia, ⁴Institut de Biomedicina de la Universitat de Barcelona (IBUB), ⁵Department of Molecular and Translational Medicine, Fibroblast Reprogramming Unit,University of Brescia

DEŞİFRE METNİ

The overall goal of this procedure is to deliver low volumes of high titer lentiviral particles carrying specific DNA constructs to the adult mouse ventricular-subventricular zone to infect all of its cell types or to the lateral ventricle to affect ependymal cells only.This method can help answer key questions about interactions that neural stem cells of the adult ventricular-subventricular niche have with their neighboring ependymal cells.The main advantage of this technique is that lentiviral vectors integrate into the genome of target cells in a cell cycle independent way, allowing for long-term diffication of rarely dividing cells.Helping me to demonstrate the procedure will be grad student Beatriz Mart

Özet

Here we describe a procedure based on the use of lentiviral particles for the long-term genetic modification of neural stem cells and/or their adjacent ependymal cells in the adult ventricular-subventricular neurogenic niche which allows the separate analysis of cell autonomous and non-autonomous, niche-dependent effects on neural stem cells.