Nonviral in situ green fluorescent protein labeling and culture of primary, adult human hair follicle epithelial progenitor cells
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Nonviral in situ green fluorescent protein labeling and culture of primary, adult human hair follicle epithelial progenitor cells. / Tiede, Stephan; Koop, Norbert; Kloepper, Jennifer E; Fässler, Reinhard; Paus, Ralf.
in: Stem cells (Dayton, Ohio), Jahrgang 27, Nr. 11, 11.2009, S. 2793-803.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Nonviral in situ green fluorescent protein labeling and culture of primary, adult human hair follicle epithelial progenitor cells
AU - Tiede, Stephan
AU - Koop, Norbert
AU - Kloepper, Jennifer E
AU - Fässler, Reinhard
AU - Paus, Ralf
PY - 2009/11
Y1 - 2009/11
N2 - In this article we show that cloning of the human K15 promoter before a green fluorescence protein (GFP)/geneticin-resistance cassette and transfection of microdissected, organ-cultured adult human scalp hair follicles generates specific K15 promoter-driven GFP expression in their stem cell-rich bulge region. K15-GFP+ cells can be visualized in situ by GFP fluorescence and 2-photon laser scanning microscopy. Vital K15-GFP+ progenitor cells can then be selected by using the criteria of their green fluorescence, adhesion to collagen type IV and fibronectin, and geneticin resistance. Propagated K15-GFP+ cells express epithelial progenitor markers, show the expected differential gene expression profile of human bulge epithelium, and form holoclones. This application of nonretroviral, K15 promoter-driven, GFP labeling to adult human hair follicles facilitates the characterization and manipulation of human epithelial stem cells, both in situ and in vitro, and should be transferable to other complex human tissues.
AB - In this article we show that cloning of the human K15 promoter before a green fluorescence protein (GFP)/geneticin-resistance cassette and transfection of microdissected, organ-cultured adult human scalp hair follicles generates specific K15 promoter-driven GFP expression in their stem cell-rich bulge region. K15-GFP+ cells can be visualized in situ by GFP fluorescence and 2-photon laser scanning microscopy. Vital K15-GFP+ progenitor cells can then be selected by using the criteria of their green fluorescence, adhesion to collagen type IV and fibronectin, and geneticin resistance. Propagated K15-GFP+ cells express epithelial progenitor markers, show the expected differential gene expression profile of human bulge epithelium, and form holoclones. This application of nonretroviral, K15 promoter-driven, GFP labeling to adult human hair follicles facilitates the characterization and manipulation of human epithelial stem cells, both in situ and in vitro, and should be transferable to other complex human tissues.
KW - Apoptosis
KW - Cell Proliferation
KW - Cell Survival
KW - Cells, Cultured
KW - Epithelial Cells
KW - Fluorescent Antibody Technique
KW - Green Fluorescent Proteins
KW - Hair Follicle
KW - Humans
KW - Immunohistochemistry
KW - Keratin-15
KW - Oligonucleotide Array Sequence Analysis
KW - Polymerase Chain Reaction
KW - Promoter Regions, Genetic
KW - Stem Cells
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1002/stem.213
DO - 10.1002/stem.213
M3 - SCORING: Journal article
C2 - 19750535
VL - 27
SP - 2793
EP - 2803
JO - STEM CELLS
JF - STEM CELLS
SN - 1066-5099
IS - 11
ER -