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Isolating Immune Cells from Mouse Embryonic Skin
Skin is the primary barrier against the external environment and develops a robust immune network for its surveillance. The origin of the resident immune cells of the skin has become a focus of interest over past a decade. Fate mapping studies have revealed that the macrophages home into the skin as early as E12.5 and are derived from the yolk sac and fetal liver. The resident γδT cells are born in the thymus and home to the skin by E16.5. Recent work from our lab has shown that the embryonic macrophages can actively remodel the extracellular matrix in skin suggesting that the skin immune system can be activated long before exposure to foreign antigens. In this chapter, we present a detailed protocol for isolating monocytes, macrophages, and epidermal dendritic T cell populations from embryonic skin.
KeywordsEmbryonic skin Isolation Macrophages Monocytes T cells Flow cytometry
We would like to thank H. Krishnamurthy and the Central Imaging and Flow Facility (CIFF) at NCBS for the use of the confocal microscopes and FACs facility. Animal work was partially supported by the National Mouse Research Resource (NaMoR) grant (BT/PR5981/MED/31/181/2012; 2013-2016) from the DBT. We thank members of the Raghavan lab for feedback. The SR lab is funded through core funds from inStem supported by the Department of Biotechnology (DBT), DBT grant (BT/PR8655/AGR/36/759/2013), and DST-SERB grant (EMR/2016/003199).
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