An improved, standardised protocol for the isolation, enrichment and targeted neural differentiation of Nestin+ progenitors from adult human dermis

Nancy Ernst, Stephan Tiede, Volker Tronnier, Charli Kruse, Christina Zechel, Ralf Paus

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Human skin-derived Nestin+ cells serve as a convenient source for autologous, adult, pluripotent progenitor cells that offer new therapeutic possibilities in cell-based regenerative medicine. However, the isolation of human Nestin+ cells has tended to be of very low efficiency and to produce highly variable cell yields. Here we report a standardised protocol that facilitates the isolation and enrichment of Nestin+ progenitor cells from enzymatically digested adult human scalp dermis. The use of distinct media like Dulbecco's modified Eagle medium supplemented with foetal bovine serum or, alternatively, serum-free, supplemented neural stem cell medium greatly affected cell morphology, proliferation and differentiation (e.g. towards a neural versus mesenchymal phenotype). Finally, Nestin+ cells were isolated from a heterogeneous dermis-derived progenitor cell population, which proliferates within clones or floating microspheres under defined serum-free culture conditions. Supplementation of the medium with epidermal growth factor and basic fibroblast growth factor as well as coating with fibronectin allowed the highest enrichment level of Nestin+ progenitors and differentiation towards neural fate. These methodological advances should greatly facilitate the isolation, culture and targeted differentiation of primary, adult human scalp skin dermis-derived Nestin+ cells.

Original languageEnglish (US)
Pages (from-to)549-555
Number of pages7
JournalExperimental dermatology
Volume19
Issue number6
DOIs
StatePublished - Jun 1 2010
Externally publishedYes

Keywords

  • Adult human skin
  • Nestin
  • Neural differentiation
  • Progenitors

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology

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