Preferential Transfection of Adult Mouse Neural Stem Cells and Their Immediate Progeny in Vivo with Polyethylenimine

Lemkine, Gregory F.; Mantero, Stefano; Migné, Carole; Raji, Aicha; Goula, Daniel; Normandie, Priscilla; Levi, Giovanni; Demeneix, Barbara

doi:10.1006/mcne.2001.1084

Cited by 41 publications

(37 citation statements)

References 27 publications

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“…Li et al(2005) reported that the highest transfection efficiency in these cells was 39.9% through lipofectamine 2000. Studies reported that the transfection efficiencies through lipofectamine 2000 or routine electroporation were lower than 20%, or sometimes even lower than 5% (Falk et al, 2002;Guo et al, 2003;Hung et al, 2004;Lemkine et al, 2002). On the other hand, the transfection efficiencies via the recombinant adenoviral or recombinant retroviral vectors were much higher.…”

Section: Discussionmentioning

confidence: 99%

Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein

Wang

Liu

et al. 2008

J. Zhejiang Univ. Sci. B

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Abstract:Objective: To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of GFP. Methods: The Ad-GFP was constructed by homologous recombination in bacteria with the AdEasy system; NSCs were isolated from rat fetal hippocampus and cultured as neurosphere suspensions. After infection with the recombinant Ad-GFP, NSCs were examined with a fluorescent microscopy and a flow cytometry for their expression of GFP. Results: After the viral infection, flow cytometry analysis revealed that the percentage of GFP-positive cells was as high as 97.05%. The infected NSCs sustained the GFP expression for above 4 weeks. After differentiated into astrocytes or neurons, they continued to express GFP efficiently. Conclusion: We have successfully constructed a viral vector Ad-GFP that can efficiently infect the primary NSCs. The reporter gene was showed fully and sustained expression in the infected cells as well as their differentiated progenies.

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Section: Discussionmentioning

confidence: 99%

Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein

Wang

Liu

et al. 2008

J. Zhejiang Univ. Sci. B

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“…1,2 The mitotically active precursor population in the adult brain is a heterogeneous population of cells that include stem cells as defined by their ability to self-renew and undergo multilineage differentiation. [3][4][5][6] In their natural state, these cells divide in the lateral ventricle and subgranular zone of the hippocampus and most eventually undergo apoptosis. 7 The mechanisms underlying adult neural stem cell apoptosis remain largely unknown despite the fact that the tremendous interest in adult neural stem cell biology is based partly upon their therapeutic potential in brain-injured states.…”

Section: Introductionmentioning

confidence: 99%

Bax limits adult neural stem cell persistence through caspase and IP3 receptor activation

2005

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Neural stem cells in the mammalian brain persist and are functional well into adulthood. There is, however, little insight into mechanisms that control adult neural stem cell survival. Mice deficient in the proapoptotic molecule Bax exhibit increased numbers of multipotent progenitor cells in the adult subventricular zone. In vitro, these progenitors behave as neural stem cells and utilize Bax and caspase activation to direct cell death. We demonstrate that the predominate mechanism underlying caspase and Bax-mediated adult neural stem cell death lies in the modulation of calcium flux through interaction with the IP3 receptor.

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“…However, such efforts are labor intensive (Haruyama et al, 2009), and gene delivery by comparison is more rapid and offers translational opportunities (Kotterman and Schaffer, 2014). Prior work in the field established some transduction of neural progenitors, but efficiency was limited and did not include Type 1 NSCs (Falk et al, 2002;Lemkine et al, 2002;van Hooijdonk et al, 2009). AAV vectors have gathered increasing momentum for basic biological investigation (Oh et al, 2014) and for clinical gene delivery (Bainbridge et al, 2008;Kotterman and Schaffer, 2014;MacLaren et al, 2014;Maguire et al, 2008Maguire et al, , 2009Nathwani et al, 2011;Ojala et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

“…Falk et al administered polyethyleneimine (PEI) complexes, containing plasmids driving reporter gene expression via enhancer elements from the second intron of the human nestin gene, to the lateral ventricle of mice and showed some selective delivery to NSCs in the SVZ, although the efficiency was limited (Falk et al, 2002). In another study, Lemkine et al used PEI-DNA complexes and showed low specificity towards mouse SVZ NSCs as compared with globular cells following delivery to the lateral ventricle (Lemkine et al, 2002). Additionally, van Hooijdonk et al (van Hooijdonk et al, 2009) used a vesicular stomatitis virus G glycoprotein-pseudotyped lentivirus to target neural progenitor cells and immature neurons in the subgranular zone of the dentate gyrus of the mouse hippocampus.…”

Section: Introductionmentioning

confidence: 99%

Enhanced selective gene delivery to neural stem cells in vivo by an adeno-associated viral variant

2015

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Neural stem cells (NSCs) are defined by their ability to self-renew and to differentiate into mature neuronal and glial cell types. NSCs are the subject of intense investigation, owing to their crucial roles in neural development and adult brain function and because they present potential targets for gene and cell replacement therapies following injury or disease. Approaches to specifically genetically perturb or modulate NSC function would be valuable for either motivation. Unfortunately, most gene delivery vectors are incapable of efficient or specific gene delivery to NSCs in vivo. Vectors based on adenoassociated virus (AAV) present a number of advantages and have proven increasingly successful in clinical trials. However, natural AAV variants are inefficient in transducing NSCs. We previously engineered a novel AAV variant (AAV r3.45) capable of efficient transduction of adult NSCs in vitro. Here, to build upon the initial promise of this variant, we investigated its in vitro and in vivo infectivity. AAV r3.45 was more selective for NSCs than mature neurons in a human embryonic stem cell-derived culture containing a mixture of cell types, including NSCs and neurons. It was capable of more efficient and selective transduction of rat and mouse NSCs in vivo than natural AAV serotypes following intracranial vector administration. Delivery of constitutively active β-catenin yielded insights into mechanisms by which this key regulator modulates NSC function, indicating that this engineered AAV variant can be harnessed for preferential modulation of adult NSCs in the hippocampus. The capacity to rapidly genetically modify these cells might greatly accelerate in vivo investigations of adult neurogenesis.

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Preferential Transfection of Adult Mouse Neural Stem Cells and Their Immediate Progeny in Vivo with Polyethylenimine

Cited by 41 publications

References 27 publications

Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein

Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein

Bax limits adult neural stem cell persistence through caspase and IP3 receptor activation

Enhanced selective gene delivery to neural stem cells in vivo by an adeno-associated viral variant

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