Name: a novel surgical approach for intratracheal administration of bioactive agents in a fetal mouse model
Uploaded: 2012-10-31T13:00:00
Description: Watch this Scientific Journal Video about A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model at JoVE.com

MC и AVdP являются научных сотрудников поддержана грантами Института продвижения инноваций по науке и технологиям во Фландрии (ВВТ-Vlaanderen). JT имеет неполный клинической стипендий исследований (Кур) от UZ Левен. DV является докторской диссертации поддержана грантом от KU Leuven, DBOF/10/062. ММДЦ является докторской диссертации поддержана грантом от Conselho Насьональ де Pesquisa электронной Desenvolvimento (CNPq) и Erasmus Mundus. Исследование было профинансировано ВВТ-Vlaanderen, ЕК грант DIMI (LSHB-CT-2005-512146) и в естественных условиях Molecular Imaging Research Group (IMIR) от KU Leuven. Мы хотели бы отметить UPenn основных векторных основана Джеймсом М. Уилсон в своем роде подарок AAV6.2 упаковки плазмиды для производства вектор rAAV.

1. Спаривание Мыши для получения нужной стадии беременности Время помощник беременных мышей NMRI так, что они 18 дней (E18) беременные (всего беременности E19.5) во время операции. До и после операции они размещены в верхней фильтр клетках при нормальной комнатной температуре и н?...

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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetoscopic+tracheal+occlusion+(FETO)+for+severe+congenital+diaphragmatic+hernia:+evolution+of+a+technique+and+preliminary+results.">Fetoscopic tracheal occlusion (FETO) for severe congenital diaphragmatic hernia: evolution of a technique and preliminary results.</a> Ultrasound Obstet. Gynecol. 24, 121-126 (2004).</li><li>Buckley, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-amniotic+delivery+of+CFTR-expressing+adenovirus+does+not+reverse+cystic+fibrosis+phenotype+in+inbred+CFTR-knockout+mice.">Intra-amniotic delivery of CFTR-expressing adenovirus does not reverse cystic fibrosis phenotype in inbred CFTR-knockout mice.</a> Mol. Ther. 16, 819-824 (2008).</li><li>Davies, L. 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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Permanent+phenotypic+correction+of+hemophilia+B+in+immunocompetent+mice+by+prenatal+gene+therapy.">Permanent phenotypic correction of hemophilia B in immunocompetent mice by prenatal gene therapy.</a> Blood. 104, 2714-2721 (2004).</li><li>Senoo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adenovirus-mediated+in+utero+gene+transfer+in+mice+and+guinea+pigs:+tissue+distribution+of+recombinant+adenovirus+determined+by+quantitative+TaqMan-polymerase+chain+reaction+assay.">Adenovirus-mediated in utero gene transfer in mice and guinea pigs: tissue distribution of recombinant adenovirus determined by quantitative TaqMan-polymerase chain reaction assay.</a> Mol. Genet. 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Критические шаги <ul><li> Мыши штамма мы решили работать только NMRI мышей, потому что они имеют обильное количество щенков (средний размер помета 14,4 ± 1,8, собственные данные), терпит вмешательства хорошо и имеют хорошие материнские характеристики. </li><li> Размещение кошелек строку через стен?...

<table border="1"><tbody><tr><td> Название реагента </td><td> Компания </td><td> Номер в каталоге </td><td> Комментарии (по желанию) </td></tr><tr><td> NMRI мышей </td><td> Janvier, Le Genest Санкт-Isle, Франция </td><td></td><td></td></tr><tr><td> Isoflurane </td><td> Isoba, Intervet / Schering-Plough Animal Health, Милтон Кейнс, Великобритания </td><td></td><td></td></tr><tr><td> Prolene 6-0 </td><td> Ethicon, Groot Bijgaarden, Бельгия </td><td></td><td></td></tr><tr><td> Викрил 5-0 </td><td> Ethicon, Groot Bijgaarden, Бельгия </td><td></td><td></td></tr><tr><td> 50 мкл Hamilton стеклянный шприц, модель 1710,5 TLLX SYR </td><td> Гамильтон, Рено, Невада, США </td><td> 5495-20 </td><td></td></tr><tr><td> Острой иглой 30G </td><td> Гамильтон, Рено, Невада, США </td><td> 7762-03 </td><td></td></tr><tr><td> 2% ксилокаин </td><td> AstraZeneca, Zoetermeer, Нидерланды </td><td></td><td></td></tr></tbody></table>

a novel surgical approach for intratracheal administration of bioactive agents in a fetal mouse model

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and acquired diseases. Apart from congenital or inherited abnormalities with the requirement for long-term expression of the delivered gene, several non-inherited perinatal conditions, where short-term gene expression or pharmacological intervention is sufficient to achieve therapeutic effects, are considered as potential future indications for this kind of approach. Candidate diseases for the application of short-term prenatal therapy could be the transient neonatal deficiency of surfactant protein B causing neonatal respiratory distress syndrome1,2 or hyperoxic injuries of the neonatal lung3. Candidate diseases for permanent therapeutic correction are Cystic Fibrosis (CF)4, genetic variants of surfactant deficiencies5 and &alpha;1-antitrypsin deficiency6.
Generally, an important advantage of prenatal gene therapy is the ability to start therapeutic intervention early in development, at or even prior to clinical manifestations in the patient, thus preventing irreparable damage to the individual. In addition, fetal organs have an increased cell proliferation rate as compared to adult organs, which could allow a more efficient gene or stem cell transfer into the fetus. Furthermore, in utero gene delivery is performed when the individual's immune system is not completely mature. Therefore, transplantation of heterologous cells or supplementation of a non-functional or absent protein with a correct version should not cause immune sensitization to the cell, vector or transgene product, which has recently been proven to be the case with both cellular and genetic therapies7.
In the present study, we investigated the potential to directly target the fetal trachea in a mouse model. This procedure is in use in larger animal models such as rabbits and sheep8, and even in a clinical setting9, but has to date not been performed before in a mouse model. When studying the potential of fetal gene therapy for genetic diseases such as CF, the mouse model is very useful as a first proof-of-concept because of the wide availability of different transgenic mouse strains, the well documented embryogenesis and fetal development, less stringent ethical regulations, short gestation and the large litter size.
Different access routes have been described to target the fetal rodent lung, including intra-amniotic injection10-12, (ultrasound-guided) intrapulmonary injection13,14 and intravenous administration into the yolk sac vessels15,16 or umbilical vein17. Our novel surgical procedure enables researchers to inject the agent of choice directly into the fetal mouse trachea which allows for a more efficient delivery to the airways than existing techniques18.

Watch this Scientific Journal Video about A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model at JoVE.com

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

Мы разработали новый подход к хирургическому интратрахеального введения биологически активных веществ в мышь плода. Маршрут доставки является более эффективным в ориентированных на легкие плода мыши, чем обычно используется внутри амниотической инъекций. Эта процедура на сегодняшний день не было описано в мышиной модели.

Новый хирургический подход к интратрахеального введения биологически активных веществ в фетальный модель мыши

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and ...

Research

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