Protocol: Adeno-Associated Virus-Mediated Gene Transfer in Ex Vivo Cultured Embryonic Mammary Gland

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http://hdl.handle.net/10138/329896

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Lan , Q & Mikkola , M L 2020 , ' Protocol: Adeno-Associated Virus-Mediated Gene Transfer in Ex Vivo Cultured Embryonic Mammary Gland ' , Journal of Mammary Gland Biology and Neoplasia , vol. 25 , no. 4 , pp. 409-416 . https://doi.org/10.1007/s10911-020-09461-4

Title: Protocol: Adeno-Associated Virus-Mediated Gene Transfer in Ex Vivo Cultured Embryonic Mammary Gland
Author: Lan, Qiang; Mikkola, Marja L.
Contributor: University of Helsinki, Institute of Biotechnology
University of Helsinki, Institute of Biotechnology
Date: 2020-12
Language: eng
Number of pages: 8
Belongs to series: Journal of Mammary Gland Biology and Neoplasia
ISSN: 1083-3021
URI: http://hdl.handle.net/10138/329896
Abstract: Branching morphogenesis of the murine mammary gland starts during late embryogenesis. It is regulated by the signals emanating both from the epithelium and the mesenchyme, yet the molecular mechanisms regulating this process remain poorly understood. We have previously developed a unique whole organ culture technique for embryonic mammary glands, which provides a powerful tool to monitor and manipulate branching morphogenesis ex vivo. Nowadays, RNA sequencing and other transcriptional profiling techniques provide robust methods to identify components of gene regulatory networks driving branching morphogenesis. However, validation of the candidate genes still mainly depends on the use of the transgenic mouse models, especially in mammary gland studies. By comparing different serotypes of recombinant adeno-associated virus (rAAVs), we found out that rAAVs provide sufficient efficiency for gene transfer with different tissue preferences depending on the serotypes of the virus. AAV-2 and AAV-8 preferentially target epithelial and mesenchymal compartments, respectively, while AAV-9 infects both tissues. Here, we describe a protocol for AAV-mediated gene transfer in ex vivo cultured murine embryonic mammary gland facilitating gene function studies on mammary gland branching morphogenesis.
Subject: 1182 Biochemistry, cell and molecular biology
Organ culture
Ex vivo
Explant culture
Adeno-associated virus
MOUSE
MORPHOGENESIS
APPENDAGE
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