Nonviral in situ green fluorescent protein labeling and culture of primary, adult human hair follicle epithelial progenitor cells

Stephan Tiede, Norbert Koop, Jennifer E. Kloepper, Reinhard Fässler, Ralf Paus

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

In this article we show that cloning of the human K15 promoter before a green fluorescence protein (GFP)/geneticinresistance cassette and transfection of microdissected, organcultured 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.

Original languageEnglish (US)
Pages (from-to)2793-2803
Number of pages11
JournalStem Cells
Volume27
Issue number11
DOIs
StatePublished - Nov 1 2009
Externally publishedYes

Fingerprint

Hair Follicle
Green Fluorescent Proteins
Stem Cells
Fluorescence
Epithelial Cells
Proteins
Collagen Type IV
Scalp
Photons
Transcriptome
Fibronectins
Confocal Microscopy
Transfection
Organism Cloning
Epithelium

Keywords

  • Adult stem cells
  • Clonal assays
  • Differentiation
  • Gene targeting
  • Green fluorescent protein
  • In vivo optical imaging
  • Progenitor cells
  • Tissue-specific stem cells

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

Cite this

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, Vol. 27, No. 11, 01.11.2009, p. 2793-2803.

Research output: Contribution to journalArticle

Tiede, Stephan ; Koop, Norbert ; Kloepper, Jennifer E. ; Fässler, Reinhard ; Paus, Ralf. / Nonviral in situ green fluorescent protein labeling and culture of primary, adult human hair follicle epithelial progenitor cells. In: Stem Cells. 2009 ; Vol. 27, No. 11. pp. 2793-2803.
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