scholarly journals Complementary In Vivo Approaches Reveal Quiescent Melanocytes in Anagen Outside the Hair Follicle Bulge

2020 ◽  
Author(s):  
Bishal Tandukar ◽  
Sandeep S. Joshi ◽  
Li Pan ◽  
Thomas J. Hornyak
Keyword(s):  
Stem Cell ◽  
Hair Follicle ◽  
Cell System ◽  
Cell Phenotype ◽  
Differentiation Markers ◽  
Outer Root Sheath ◽  
Root Sheath ◽  
Melanocyte Stem Cells ◽  
Stem Cell System

ABSTRACTMelanocyte stem cells (McSCs) are key components of the hair follicle (HF) stem cell system that are derived from neural crest during embryogenesis and are responsible for regeneration of differentiated melanocytes during successive HF cycles. Our previous research has shown presence of two subsets of phenotypically and functionally distinct McSCs exist in murine telogen HFs, CD34+ McSCs in the bulge/lower permanent portion (LPP) and CD34− McSCs in the secondary hair germ (SHG). Whether these subsets are maintained independently or exist in a developmental hierarchy is not yet known. Using Dct-H2BGFP mice, we analyzed the quiescent and proliferative properties of McSCs and melanocytes in anagen and telogen. We found unexpectedly that Kit+Nestin− quiescent melanocytes are maintained outside of the bulge/LPP region throughout anagen in addition to the Kit+Nestin+ quiescent melanocytes of the bulge/LPP. Both subpopulations express lower levels of melanocyte differentiation markers Mitf, Pax3, Dct, Tyrp1 and Tyr compared to differentiated melanocytes of the HF bulb/matrix. These results suggest that quiescent melanocytes localized in the outer root sheath, both in and below the bulge/LPP) retain the stem cell phenotype observed in quiescent McSCs during telogen. This finding has implications for maintenance of distinct subsets of McSCs throughout successive HF cycles.

scholarly journals CCN2 modulates hair follicle cycling in mice

2013 ◽  
Vol 24 (24) ◽  
pp. 3939-3944 ◽  
Author(s):  
Shangxi Liu ◽  
Andrew Leask
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Hair Follicle ◽  
Cell Activity ◽  
Adult Life ◽  
Tissue Growth ◽  
Hair Follicles ◽  
Outer Root Sheath ◽  
Root Sheath ◽  
Dermal Papillae

It is critical to understand how stem cell activity is regulated during regeneration. Hair follicles constitute an important model for organ regeneration because, throughout adult life, they undergo cyclical regeneration. Hair follicle stem cells—epithelial cells located in the follicle bulge—are activated by periodic β-catenin activity, which is regulated not only by epithelial-derived Wnt, but also, through as-yet-undefined mechanisms, the surrounding dermal microenvironment. The matricellular protein connective tissue growth factor (CCN2) is secreted into the microenvironment and acts as a multifunctional signaling modifier. In adult skin, CCN2 is largely absent but is unexpectedly restricted to the dermal papillae and outer root sheath. Deletion of CCN2 in dermal papillae and the outer root sheath results in a shortened telogen-phase length and elevated number of hair follicles. Recombinant CCN2 causes decreased β-catenin stability in keratinocytes. In vivo, loss of CCN2 results in elevated numbers of K15-positive epidermal stem cells that possess elevated β-catenin levels and β-catenin–dependent reporter gene expression. These results indicate that CCN2 expression by dermal papillae cells is a physiologically relevant suppressor of hair follicle formation by destabilization of β-catenin and suggest that CCN2 normally acts to maintain stem cell quiescence.

scholarly journals PSF1 Is Essential for Early Embryogenesis in Mice

2005 ◽  
Vol 25 (23) ◽  
pp. 10528-10532 ◽  
Author(s):  
Masaya Ueno ◽  
Machiko Itoh ◽  
Lingyu Kong ◽  
Kazushi Sugihara ◽  
Masahide Asano ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell System ◽  
Early Embryogenesis ◽  
Hematopoietic Stem ◽  
Egg Extracts ◽  
A Cell ◽  
Stem Cell System ◽  
Initiation Of Dna Replication ◽  
Adult Bone

ABSTRACT Psf1 (partner of sld five 1) forms a novel heterotetramer complex, GINS (Go, Ichi, Nii, and San; five, one, two, and three, respectively, in Japanese), with Sld5, Psf2, and Psf3. The formation of this complex is essential for the initiation of DNA replication in yeast and Xenopus laevis egg extracts. Although all of the components are well conserved in higher eukaryotes, the biological function in vivo is largely unknown. We originally cloned the mouse ortholog of PSF1 from a hematopoietic stem cell cDNA library and found that PSF1 is expressed in blastocysts, adult bone marrow, and testis, in which the stem cell system is active. Here we used the gene-targeting technique to determine the physiological function of PSF1 in vivo. Mice homozygous for a nonfunctional mutant of PSF1 died in utero around the time of implantation. PSF1 − / − blastocysts failed to show outgrowth in culture and exhibited a cell proliferation defect. Our data clearly indicate that PSF1 is required for early embryogenesis.

scholarly journals Migration of Melanoblasts into the Developing Murine Hair Follicle Is Accompanied by Transient c-Kit Expression

2002 ◽  
Vol 50 (6) ◽  
pp. 751-766 ◽  
Author(s):  
Eva M. J. Peters ◽  
Desmond J. Tobin ◽  
Natasha Botchkareva ◽  
Marcus Maurer ◽  
Ralf Paus
Keyword(s):  
Stem Cell ◽  
Hair Follicle ◽  
Stem Cell Factor ◽  
Mouse Skin ◽  
Dermal Papilla ◽  
Hair Follicles ◽  
Cell Populations ◽  
Outer Root Sheath ◽  
Hair Bulb ◽  
Root Sheath

Disruption of the c-Kit/stem cell factor (SCF) signaling pathway interferes with the survival, migration, and differentiation of melanocytes during generation of the hair follicle pigmentary unit. We examined c-Kit, SCF, and S100 (a marker for precursor melanocytic cells) expression, as well as melanoblast/melanocyte ultrastructure, in perinatal C57BL/6 mouse skin. Before the onset of hair bulb melanogenesis (i.e., stages 0–4 of hair follicle morphogenesis), strong c-Kit immunoreactivity (IR) was seen in selected non-mela-nogenic cells in the developing hair placode and hair plug. Many of these cells were S100-IR and were ultrastructurally identified as melanoblasts with migratory appearance. During the subsequent stages (5 and 6), increasingly dendritic c-Kit-IR cells successively invaded the hair bulb, while S100-IR gradually disappeared from these cells. Towards the completion of hair follicle morphogenesis (stages 7 and 8), several distinct follicular melanocytic cell populations could be defined and consisted broadly of (a) undifferentiated, non-pigmented c-Kit-negative melanoblasts in the outer root sheath and bulge and (b) highly differentiated melanocytes adjacent to the hair follicle dermal papilla above Auber's line. Widespread epithelial SCF-IR was seen throughout hair follicle morphogenesis. These findings suggest that melanoblasts express c-Kit as a prerequisite for migration into the SCF-supplying hair follicle epithelium. In addition, differentiated c-Kit-IR melanocytes target the bulb, while non-c-Kit-IR melanoblasts invade the outer root sheath and bulge in fully developed hair follicles.

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

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 154
Author(s):  
Hanluo Li ◽  
Federica Francesca Masieri ◽  
Marie Schneider ◽  
Alexander Bartella ◽  
Sebastian Gaus ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Hair Follicle ◽  
Cell Populations ◽  
Stem Cell Markers ◽  
Outer Root Sheath ◽  
Root Sheath ◽  
Gene And Protein Expression ◽  
Stem Cell Populations

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.

The in vivo effect of chelidonine on the stem cell system of planarians

2012 ◽  
Vol 686 (1-3) ◽  
pp. 1-7 ◽  
Author(s):  
Maria Emilia Isolani ◽  
Daniele Pietra ◽  
Linda Balestrini ◽  
Alice Borghini ◽  
Paolo Deri ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell System ◽  
Stem Cell System

Functional Melanocytes Obtained From Rabbit Hair Follicle Outer Root Sheath

2020 ◽  
Author(s):  
Hanluo Li ◽  
Christina-Marie Baumbach ◽  
Marie Schneider ◽  
Katrin Schwalenberg ◽  
Tina Kottek ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hair Follicle ◽  
Ex Vivo ◽  
Nile Blue ◽  
Experimental Models ◽  
Hair Follicles ◽  
Blue Staining ◽  
Outer Root Sheath ◽  
Root Sheath

Abstract BackgroundMelanocytes have been cultivated from the outer root sheath of hair follicles for two decades. So far, these isolation and culturing procedures have been carried out from human and mouse follicles. In this study, we have translated the established procedure for obtaining melanocytes from stem cells and precursors of human hair follicle to a rabbit species in order to isolate and cultivate rabbit melanocytes from whisker follicle outer root sheath (ORS), hereby named rMORS, and compare them to rabbit epidermal rabbit melanocytes (rEMs).ResultsThe rMORS were isolated and cultured by extracting whisker follicles from enzymatically digested skin and submitting them to the air-liquid interface hypoxic conditions. The cells were allowed to migrate from the follicle ORS onto the nylon mesh of Transwell inserts, collected and subcultured in melanocyte culture medium for 11 passages. From early passages on, the rMORS cells displayed typical melanocyte characteristics comparable to those of rEM used as experimental control. Melanocyte features were assessed on morphological level by the means of microscopy, functional and gene expression level. Functionality was qualitatively assessed by the Von Kossa and Nile Blue staining of melanin and by biochemically quantifying melanin content and Tyrosinase activity. Gene expression of neuroectodermal and melanocytic lineage markers NES, PAX3, MITF, TYR, CKIT and PMEL was determined by the means of qRT-PCR. ConclusionsWe concluded that the method of isolating and culturing rMORS efficiently and reproducibly corresponded to the prior art method established in human follicle, yielding melanocytes comparable to the rabbit epidermal melanocytes in all assessed features. This method is not crucial in terms of regenerative therapy, it rather paves way for a non-invasive ex vivo sampling of hair and it may give way to deeper insights into rabbit follicle biology or to postulating useful species-specific in vivo experimental models.

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