The Middle Part of the Plucked Hair Follicle Outer Root Sheath Is Identified as an Area Rich in Lineage-Specific Stem Cell Markers.

Hanluo Li,Ulrich Sack,Jan-Christoph Simon,Federica Francesca Masieri,Danijela Maksimovic-Ivanic,Alexander Bartella,Sebastian Hahnel,Rüdiger Zimmerer,Marie Schneider,Vuk Savkovic,Sanja Mijatovic,Bernd Lethaus,Sebastian Gaus

doi:10.3390/biom11020154

Hanluo Li, Ulrich Sack + Show 11 more

Open Access

https://doi.org/10.3390/biom11020154

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Abstract

Hair follicle outer root sheath (ORS) is a putative source of stem cells with therapeutic capacity. ORS contains several multipotent stem cell populations, primarily in the distal compartment of the bulge region. However, the bulge is routinely obtained using invasive isolation methods, which require human scalp tissue ex vivo. Non-invasive sampling has been standardized by means of the plucking procedure, enabling to reproducibly obtain the mid-ORS part. The mid-ORS shows potential for giving rise to multiple stem cell populations in vitro. To demonstrate the phenotypic features of distal, middle, and proximal ORS parts, gene and protein expression profiles were studied in physically separated portions. The mid-part of the ORS showed a comparable or higher NGFR, nestin/NES, CD34, CD73, CD44, CD133, CK5, PAX3, MITF, and PMEL expression on both protein and gene levels, when compared to the distal ORS part. Distinct subpopulations of cells exhibiting small and round morphology were characterized with flow cytometry as simultaneously expressing CD73/CD271, CD49f/CD105, nestin, and not CK10. Potentially, these distinct subpopulations can give rise to cultured neuroectodermal and mesenchymal stem cell populations in vitro. In conclusion, the mid part of the ORS holds the potential for yielding multiple stem cells, in particular mesenchymal stem cells.

Highlights

Washed hair follicles were further prepared for purposes of histological characterization, flow cytometry (FCM), and qRT-PCR
To assess the anatomic structure of the human hair follicle outer root sheath (ORS), marker- protein expression profiles were visualized in histological sections of plucked human anagen hair follicles
The bulge part at the distal region of the ORS was lost in the process of plucking (Figure 1A, Hematoxylin & Eosin (H&E)) and its remains were not carried over with the rest of the plucked follicle

Summary

Introduction

It is very versatile in terms of in vitro scalable cell cultivation and multi-lineage differentiation. Other group of multipotent stem cells have been identified in the interfollicular epidermis [9] and isthmus region [10] of the hair follicle, characterized by the expression of the transmembrane receptor Lgr. Other group of multipotent stem cells have been identified in the interfollicular epidermis [9] and isthmus region [10] of the hair follicle, characterized by the expression of the transmembrane receptor Lgr6 Due to their highly naïve developmental status and the ability to generate all cell lineages of the skin [11], Lgr6+ cells are named “mother of skin cells” [10]. To the neural and neuroectodermal/skin stem cell lineages, the mesenchymal stem cell (MSC)

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