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Published on: January 30, 2021
Generation and characterization of multipotent stem cells from established dermal cultures
Rebecca P Hill1, Karl Gledhill, Aaron Gardner
1School of Biological and Biomedical Sciences, Durham University, Durham, County Durham, United Kingdom.
Plos One
|December 11, 2012
Summary
Adult human dermal fibroblasts (DF) cultured in monolayers can generate skin-derived precursor cells (SKPs), termed m-SKPs. This method offers a scalable source of m-SKPs, which retain multipotency and can be cryopreserved.
Area of Science:
- Stem Cell Biology
- Dermatology
- Regenerative Medicine
Background:
- Human multipotent skin-derived precursor cells (SKPs) are crucial for skin regeneration but traditionally sourced from limited donor tissues.
- Adult human dermal fibroblasts (DF) represent a more accessible cell population.
Purpose of the Study:
- To investigate the potential of pre-cultured adult human dermal fibroblasts (DF) to form multipotent skin-derived precursor cells (m-SKPs).
- To characterize the properties and differentiation potential of these novel m-SKPs.
- To explore factors influencing m-SKP yield and compare with existing SKP sources.
Main Methods:
- Adult human dermal fibroblasts (DF) from normal and diseased skin were cultured in monolayers.
- Formation and passaging of m-SKPs were assessed.
- Protein and transcriptional markers (nestin, fibronectin, versican, p75NTR, Nanog, Dermo-1) were analyzed.
- Differentiation potential into mesenchymal and neural lineages was induced and evaluated.
- m-SKP yield was correlated with donor age, αSMA expression, and anatomical origin.
Main Results:
- Normal and diseased adult human DF in monolayers successfully formed multipotent SKPs (m-SKPs).
- m-SKPs expressed key SKP, neural crest, embryonic, and mesenchymal stem cell markers.
- m-SKPs demonstrated multipotency, differentiating into both mesenchymal and neural lineages.
- Cryopreservation of original DF cultures maintained m-SKP yield, while extensive passaging reduced it.
- Neonatal foreskin yielded more m-SKPs but had reduced capacity after passaging; adult DF with higher αSMA expression yielded more m-SKPs.
- Human hair follicle dermal papilla cells in monolayers unexpectedly failed to form m-SKPs.
Conclusions:
- Expanded dermal fibroblast (DF) cultures provide a viable and potentially scalable source for generating multipotent skin-derived precursor cells (m-SKPs).
- m-SKPs exhibit characteristics and differentiation potential similar to traditionally sourced SKPs.
- Skin-derived precursor cell generation capacity varies with donor age, anatomical site, and culture conditions, highlighting heterogeneity.
- The progenitor status of human hair follicle dermal papilla cells warrants further investigation, as monolayer cultures did not yield m-SKPs.
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