In vitro identification of a stem cell population from canine hair follicle bulge region

Abstract

Skin is an extensive and easily accessible organ possessing various cell types that are constantly renewed. Previous studies have suggested the presence of a stem cell niche at the bulge region of the hair follicle, which contains cells positive for CD200 and CD34. Thus, this study sought to identify these cell populations in canine skin cells using the following methods 1- collecting samples of adult and fetal skin and isolating and culturing these cells using a method of simple enzymatic digestion and 2- testing the cell cultures for CD200 and CD34 in vitro and comparing them with skin tissue samples (in situ). Immunofluorescence results were negative for both CD200 and CD34 in frozen and paraffin embedded tissue, whereas the analysis showed that cultured cells positive for CD34, CD200 and double positive cells could be visualized in different percentages. Additionally, the pluripotency marker OCT4 was positive in the isolated cells. Analysis of CD34, CD200 and OCT4 by RT-qPCR showed that there is expression in fetal and adult cells, although no difference was observed between groups. Our results suggest that bulge stem cells from both fetuses and adult dogs were reported with the use of CD34 and CD200 markers in this study, and further techniques for cell isolation and in vitro cultivation are needed in order to obtain enriched populations of skin stem cells in dogs.

ADSCs inhibit photoaging‐ and photocarcinogenesis‐related inflammatory responses and extracellular matrix degradation

“…We analyzed α6 integrin and CD34 expression in skin tissue in the three groups because the positive expression of these markers indicates HFSCs. 18,23,24 Immunofluorescence results showed that UVB exposure downregulated CD34 and α6 integrin expression, which was increased after ADSCs transplantation. These findings suggest that the quantity of HFSCs decreased in photoaging skin and increased after ADSCs transplantation.…”

“…Wang transferred a chamber containing cultured HFSCs with positive expression of α6‐integrin and CD34, as identified by FACS analysis, through a hole in clipped skin, and hair follicles formed 21 days later. We analyzed α6 integrin and CD34 expression in skin tissue in the three groups because the positive expression of these markers indicates HFSCs . Immunofluorescence results showed that UVB exposure downregulated CD34 and α6 integrin expression, which was increased after ADSCs transplantation.…”

3D skin models in domestic animals

“…Different stem cells with have also been identified in the upper part of the hair follicle (isthmus and junction zone) [57]. Stem cells of the follicle hair bulge region have been identified in different mammalian species: dog [58][59][60], sheep [61], and pig [62]. In birds, epithelial stem cells are located in the collar bulge of the feather follicle above the dermal papilla [63].…”

Transcriptome Profiling and Differential Gene Expression in Canine Microdissected Anagen and Telogen Hair Follicles and Interfollicular Epidermis

“…The renewal of the IFE and the HF relies on stem cells and their progenitors. Several stem cell markers, such as LGR5, CD34, SOX9 and KRT15 (HF bulge stem cells markers) [ 9 , 10 , 11 , 12 , 13 ] or LGR-6 (expressed in the upper HF, sebaceous gland and IFE) have been described in humans and rodents [ 14 , 15 ] and their expression has also been investigated in dogs [ 10 , 16 , 17 , 18 ]. Stem cell quiescence and activation, as well as the subsequent events leading to epidermal differentiation and the maintenance of the hair cycle, are tightly regulated and involve numerous signals derived from epithelial, neuroendocrine and mesenchymal cells [ 19 , 20 , 21 ].…”