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.
Embryonic skin Isolation Macrophages Monocytes T cells Flow cytometry
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
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).
Nestle FO, Di Meglio P et al (2009) Skin immune sentinels in health and disease. Nat Rev Immunol 9(10):679–679CrossRefGoogle Scholar
Heath WR, Carbone FR (2013) The skin-resident and migratory immune system in steady state and memory: innate lymphocytes, dendritic cells and T cells. Nat Immunol 14(10):978–985CrossRefGoogle Scholar
Jiang X et al (2010) Embryonic trafficking of gammadelta T cells to skin is dependent on E/P selectin ligands and CCR4. Proc Natl Acad Sci U S A 107(16):7443–7448CrossRefGoogle Scholar
Gurish MF, Austen KF (2012) Developmental origin and functional specialization of mast cell subsets. Immunity 37(1):25–33CrossRefGoogle Scholar
Hoeffel G et al (2012) Adult Langerhans cells derive predominantly from embryonic fetal liver monocytes with a minor contribution of yolk sac-derived macrophages. J Exp Med 209(6):1167–1181CrossRefGoogle Scholar
Castellana D et al (2014) Macrophages contribute to the cyclic activation of adult hair follicle stem cells. PLoS Biol 12(12):e1002002CrossRefGoogle Scholar