TY - CHAP
T1 - Isolation and characterization of cutaneous epithelial stem cells
AU - Gillespie, Stephanie R.
AU - Owens, David M.
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2018.
PY - 2019
Y1 - 2019
N2 - The outer layer of mammalian skin is a multilayered epithelium that perpetually renews multiple differentiated lineages. During homeostasis, the maintenance of skin epithelial turnover is ensured by regionalized populations of stem cells that largely remain dedicated to distinct epithelial lineages including squamous, follicular, sebaceous, Merkel, and sweat glands. Cutting edge developments in this field have focused on: (1) stem cell activation cues derived from a number of extrinsic sources including neurons, dermal fibroblasts and adipocyte, and immune cells; and (2) characterization of epithelial stem cell homeostasis via hierarchical versus stochastic paradigms. The techniques outlined in this chapter are designed to facilitate such studies and describe basic procedures for cutaneous stem cell isolation and purification, which are based on leveraging their unique expression of surface proteins for simultaneous targeting and purifying of multiple subpopulations in adult skin. In addition, protocols for assessment of in vitro and ex vivo progenitor capacity as well as techniques to visualize progenitor populations in whole skin are discussed.
AB - The outer layer of mammalian skin is a multilayered epithelium that perpetually renews multiple differentiated lineages. During homeostasis, the maintenance of skin epithelial turnover is ensured by regionalized populations of stem cells that largely remain dedicated to distinct epithelial lineages including squamous, follicular, sebaceous, Merkel, and sweat glands. Cutting edge developments in this field have focused on: (1) stem cell activation cues derived from a number of extrinsic sources including neurons, dermal fibroblasts and adipocyte, and immune cells; and (2) characterization of epithelial stem cell homeostasis via hierarchical versus stochastic paradigms. The techniques outlined in this chapter are designed to facilitate such studies and describe basic procedures for cutaneous stem cell isolation and purification, which are based on leveraging their unique expression of surface proteins for simultaneous targeting and purifying of multiple subpopulations in adult skin. In addition, protocols for assessment of in vitro and ex vivo progenitor capacity as well as techniques to visualize progenitor populations in whole skin are discussed.
KW - Epithelial lineage
KW - Epithelial progenitor markers
KW - Epithelial stem cell homeostasis
KW - Skin differentiation
KW - Skin reconstitution assay
UR - http://www.scopus.com/inward/record.url?scp=85060632962&partnerID=8YFLogxK
U2 - 10.1007/7651_2018_171
DO - 10.1007/7651_2018_171
M3 - Chapter
C2 - 30027508
AN - SCOPUS:85060632962
T3 - Methods in Molecular Biology
SP - 87
EP - 99
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -