Faculty of Veterinary Medicine, Institute of Anatomy, Histology and Embryology, Leipzig University, Leipzig, Germany.
Saxon Incubator for Clinical Translation, Leipzig University, Leipzig, Germany.
Cytometry A. 2018 Jan;93(1):104-114. doi: 10.1002/cyto.a.23198. Epub 2017 Sep 2.
The easily accessible niche represented by skin and its appendages may serve as a promising source to complement modern regenerative medicine for horses. In humans and in animal models for human medicine, the hair follicle and its stem cell niches are well characterized. Since literature in this field of equine research is scarce, we sought to analyze cells of the dermal stem cell niche of the equine hair follicle morphologically and for a subset of markers useful for cell characterization via immunolabeling. We cultured equine forelock skin explants to obtain cultures with cells migrating from the hair follicles. Isolation of cells revealed typical fibroblast morphology with a strong tendency to aggregate and form spheroids. For immunofluorescent characterization of primary isolations, we tested an antibody panel consisting of lineage makers for the dermal compartment of the hair follicle, markers associated with an undifferentiated cell status and markers for epithelial cell types as negative controls. All antibodies used were also tested on equine skin sections. The isolated cells displayed clear profiles of dermal and undifferentiated cells. To substantiate our findings, we tested our primary isolations for established equine multipotent mesenchymal stromal cell antigen expression markers in flow cytometry experiments yielding strong convergence. The data presented here provide insights to a stem cell source in horses almost unnoticed to date. The basic investigations of the equine dermal hair follicle stem cell niche confirm the expression of standard markers used in other species and lay the foundation for future studies on this easily available adult stem cell source. © 2017 International Society for Advancement of Cytometry.
皮肤及其附属物的易接近小生境可能成为补充现代马再生医学的有前途的来源。在人类和用于人类医学的动物模型中,毛囊及其干细胞小生境得到了很好的描述。由于该领域的马研究文献很少,我们试图通过免疫标记分析马毛囊的真皮干细胞小生境的细胞,从形态上和用于细胞特征鉴定的一组标记物进行分析。我们培养马额发皮肤外植体以获得从毛囊迁移而来的细胞培养物。细胞分离显示出典型的成纤维细胞形态,具有强烈的聚集和形成球体的趋势。为了对原代分离物进行免疫荧光特征分析,我们测试了一个由毛囊真皮区谱系标志物、未分化细胞状态相关标志物和上皮细胞类型标志物组成的抗体组合,作为阴性对照。所有使用的抗体也在马皮肤切片上进行了测试。分离的细胞显示出清晰的真皮和未分化细胞特征。为了证实我们的发现,我们在流式细胞术实验中测试了我们的原代分离物中已建立的马多能间充质基质细胞抗原表达标志物,得到了强烈的收敛。这里提供的数据分析为迄今为止几乎未被注意到的马的干细胞来源提供了新的认识。对马真皮毛囊干细胞小生境的基础研究证实了在其他物种中使用的标准标记物的表达,并为该易获得的成体干细胞来源的未来研究奠定了基础。