Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration

Mort, Richard L. and Keighren, Margaret and Hay, Leonard and Jackson, Ian J. (2014) Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration. Journal of Visualized Experiments (87). ISSN 1940-087X

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Abstract

Melanoblasts are the neural crest derived precursors of melanocytes; the cells responsible for producing the pigment in skin and hair. Melanoblasts migrate through the epidermis of the embryo where they subsequently colonize the developing hair follicles(1,2). Neural crest cell migration is extensively studied in vitro but in vivo methods are still not well developed, especially in mammalian systems. One alternative is to use ex vivo organotypic culture(3-6). Culture of mouse embryonic skin requires the maintenance of an air-liquid interface (ALI) across the surface of the tissue(3,6). High resolution live-imaging of mouse embryonic skin has been hampered by the lack of a good method that not only maintains this ALI but also allows the culture to be inverted and therefore compatible with short working distance objective lenses and most confocal microscopes. This article describes recent improvements to a method that uses a gas permeable membrane to overcome these problems and allow high-resolution confocal imaging of embryonic skin in ex vivo culture(6). By using a melanoblast specific Cre-recombinase expressing mouse line combined with the R26YFPR reporter line we are able to fluorescently label the melanoblast population within these skin cultures. The technique allows live-imaging of melanoblasts and observation of their behavior and interactions with the tissue in which they develop. Representative results are included to demonstrate the capability to live-image 6 cultures in parallel.

Item Type:
Journal Article
Journal or Publication Title:
Journal of Visualized Experiments
Uncontrolled Keywords:
/dk/atira/pure/subjectarea/asjc/2800
Subjects:
ID Code:
84128
Deposited By:
Deposited On:
20 Jan 2017 13:14
Refereed?:
Yes
Published?:
Published
Last Modified:
16 Nov 2020 07:14