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

Research output: Contribution to journalJournal article

Published

Standard

Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration. / Mort, Richard L.; Keighren, Margaret; Hay, Leonard; Jackson, Ian J.

In: Journal of Visualized Experiments, No. 87, 19.05.2014.

Research output: Contribution to journalJournal article

Harvard

Mort, RL, Keighren, M, Hay, L & Jackson, IJ 2014, 'Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration', Journal of Visualized Experiments, no. 87. https://doi.org/10.3791/51352

APA

Mort, R. L., Keighren, M., Hay, L., & Jackson, I. J. (2014). Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration. Journal of Visualized Experiments, (87). https://doi.org/10.3791/51352

Vancouver

Mort RL, Keighren M, Hay L, Jackson IJ. Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration. Journal of Visualized Experiments. 2014 May 19;(87). https://doi.org/10.3791/51352

Author

Mort, Richard L. ; Keighren, Margaret ; Hay, Leonard ; Jackson, Ian J. / Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration. In: Journal of Visualized Experiments. 2014 ; No. 87.

Bibtex

@article{986e19d395194e2ebe0e2f5f6ab4853d,
title = "Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration",
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.",
keywords = "Animals, Cell Movement, Female, Male, Melanocytes, Mice, Microscopy, Confocal, Organ Culture Techniques, Skin, Journal Article, Research Support, Non-U.S. Gov't, Video-Audio Media",
author = "Mort, {Richard L.} and Margaret Keighren and Leonard Hay and Jackson, {Ian J.}",
year = "2014",
month = may
day = "19",
doi = "10.3791/51352",
language = "English",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "87",

}

RIS

TY - JOUR

T1 - Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration

AU - Mort, Richard L.

AU - Keighren, Margaret

AU - Hay, Leonard

AU - Jackson, Ian J.

PY - 2014/5/19

Y1 - 2014/5/19

N2 - 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.

AB - 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.

KW - Animals

KW - Cell Movement

KW - Female

KW - Male

KW - Melanocytes

KW - Mice

KW - Microscopy, Confocal

KW - Organ Culture Techniques

KW - Skin

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

KW - Video-Audio Media

U2 - 10.3791/51352

DO - 10.3791/51352

M3 - Journal article

C2 - 24894489

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 87

ER -