Equine Hoof Stem Progenitor Cells (HPC) CD29 + /Nestin + /K15 + – a Novel Dermal/epidermal Stem Cell Population With a Potential Critical Role for Laminitis Treatment

Abstract

Laminitis is a life threating, extremely painful and frequently recurrent disease of horses which affects hoof structure. It results from the disruption of blood flow to the laminae, contributing to laminitis and in severe separation of bone from the hoof capsule. Still, the pathophysiology of the disease remains unclear, mainly due to its complexity. In the light of the presented data, in the extremally difficult process of tissue structure restoration after disruption, a novel type of progenitor cells may be involved. Herein, we isolated and performed the initial characterization of stem progenitor cells isolated from the coronary corium of the equine feet (HPC). Phenotype of the cells was investigated with flow cytometry and RT-qPCR revealing the presence of nestin, CD29, and expression of progenitor cell markers including SOX2, OCT4, NANOG and K14. Morphology of HPC was investigated with light, confocal and SEM microscopes. Cultured cells were characterised by spindle shaped morphology, eccentric nuclei, elongated mitochondria, and high proliferation rate. Plasticity and multilineage differentiation potential was confirmed by specific staining and gene expression analysis. We conclude that HPC exhibit in vitro expansion and plasticity similar to mesenchymal stem cells, which can be isolated from the equine foot, and may be directly involved in the pathogenesis and recovery of laminitis. Obtained results are of importance to the field of laminitis treatment as determining the repairing cell populations could contribute to the discovery of novel therapeutic targets and agents including and cell‐based therapies for affected animals.

Obesity affects mitochondrial metabolism and proliferative potential of equine endometrial progenitor cells

“…Documentation of cultures was performed using an inverted microscope (Axio Observer A.1, Zeiss, Oberkochen, Germany) with Power Shot digital camera (Canon, Woodhatch, UK). Images were processed and analyzed, as was previously described [21].…”

“…The analysis was performed using FACSLyric™ Clinical Flow Cytometry System (Becton-Dickinson and Company, Franklin Lakes, NJ, USA). The detailed staining protocol was described elsewhere [21], while antibodies used for analysis are listed in Table 4S.…”

Stem Cell Therapeutics in veterinary medicine in India

“…MSCs therapeutic effect in musculoskeletal ailments like dog, rabbit and horse osteoarthritis/ cartilage defects (Kriston-Pál et al 2017, Broeckx et al 2019, Gugjoo et al 2019, Huňáková et al 2020, dog bone affections (Tsuzuki et al 2014, Song et al 2017, dog periodontal defects (Takewaki et al 2017), dog and horse muscle/ tendon tears/ injuries (Beerts et al 2017;Gibson et al 2017, Taroni et al 2017, Depuydt et al 2021) and dog myocardial affections (Pogue et al 2013, Yang et al 2021) have been evaluated. Under nonmusculoskeletal tissue ailments, MSCs applications in dog, cattle and horse skin/teat/ corneal/vocal folds injuries/ ulcers (Madhu et al 2014, Zubin et al 2015, Iacono et al 2016, Iravani et al 2017, Khashjoori et al 2019, Mund et al 2020, dog keratoconjunctivitis sicca (KCS) (Villatoro et al 2015, Bittencourt et al 2016, dog spinal cord injuries (SCI) (Escalhão et al 2017, Bhat et al 2019, Sharun et al 2021, dog sciatic nerve injuries (Ding et al 2010), dog meningoencephalomyelitis of unknown origin (MUO) (Zeira et al 2015), dog hepatocutaneous syndrome (Nam et al 2017), dog and cat acute kidney injury (Lee et al 2017b), dog intervertebral disc diseases (Lee et al 2009, Sharun et al 2020 (Ferrer et al 2016) and equine laminitis (Marycz et al 2021). Recently, extended evaluation to canine diabetes, liver affections, tumors, etc have also been started .…”

A Comparative Study of Equine Hoof Progenitor Cells and Adipose-Derived Stem Cells in Hyperinsulinemia

“…Furthermore, numerous soluble mediators, inflammatory cytokines, and trophic and growth factors, including vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), brain-derived neurotrophic factor (BDNF), insulin-like growth factor (IGF), basic fibroblast growth factor (bFGF), transforming growth factor beta 1 (TGF-1) and interleukins, as well as paracrine factors that aid tissue regeneration, wound healing, and organ regeneration, are secreted by ASC [14]. Recently, we isolated and described new stem progenitor cells (HPC) that reside in the coronary corium of the equine hoof [15]. These cells have shown high levels of gene expression of surface markers such as cell adhesion molecules CD29 and an immunoglobulin superfamily membrane protein CD200, as well as nestin, a class VI intermediate filament protein, keratin 15 (K15), vascular endothelial growth factor (VEGF), and angiopoietin-1 (Ang1), indicating their high potential role in regeneration of the dermis and epidermis during laminitis [15].…”

“…Recently, we isolated and described new stem progenitor cells (HPC) that reside in the coronary corium of the equine hoof [15]. These cells have shown high levels of gene expression of surface markers such as cell adhesion molecules CD29 and an immunoglobulin superfamily membrane protein CD200, as well as nestin, a class VI intermediate filament protein, keratin 15 (K15), vascular endothelial growth factor (VEGF), and angiopoietin-1 (Ang1), indicating their high potential role in regeneration of the dermis and epidermis during laminitis [15]. Furthermore, as with other stem cell populations, HPC may be affected by inflammation in the same way that ASC are affected by equine metabolic syndrome (EMS), a disorder in which inflammation is one of the critical components for the development of insulin resistance [16,17].…”