An improved, standardised protocol for the isolation, enrichment and targeted neural differentiation of Nestin+ progenitors from adult human dermis
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An improved, standardised protocol for the isolation, enrichment and targeted neural differentiation of Nestin+ progenitors from adult human dermis. / Ernst, Nancy; Tiede, Stephan; Tronnier, Volker; Kruse, Charli; Zechel, Christina; Paus, Ralf.
In: EXP DERMATOL, Vol. 19, No. 6, 06.2010, p. 549-55.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - An improved, standardised protocol for the isolation, enrichment and targeted neural differentiation of Nestin+ progenitors from adult human dermis
AU - Ernst, Nancy
AU - Tiede, Stephan
AU - Tronnier, Volker
AU - Kruse, Charli
AU - Zechel, Christina
AU - Paus, Ralf
PY - 2010/6
Y1 - 2010/6
N2 - 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.
AB - 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.
KW - Actins
KW - Adult
KW - Adult Stem Cells
KW - Aged
KW - Cell Culture Techniques
KW - Cell Differentiation
KW - Cell Proliferation
KW - Cell Separation
KW - Culture Media
KW - Culture Media, Serum-Free
KW - Dermis
KW - Female
KW - Fibronectins
KW - Gene Expression
KW - Humans
KW - Intermediate Filament Proteins
KW - Male
KW - Mesoderm
KW - Middle Aged
KW - Nerve Tissue Proteins
KW - Nestin
KW - Neurofilament Proteins
KW - Neurons
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1111/j.1600-0625.2009.01041.x
DO - 10.1111/j.1600-0625.2009.01041.x
M3 - SCORING: Journal article
C2 - 20100195
VL - 19
SP - 549
EP - 555
JO - EXP DERMATOL
JF - EXP DERMATOL
SN - 0906-6705
IS - 6
ER -