Equine Peripheral Blood-Derived Progenitors in Comparison to Bone Marrow-Derived Mesenchymal Stem Cells

Abstract

Fibroblast-like cells isolated from peripheral blood of human, canine, guinea pig, and rat have been demonstrated to possess the capacity to differentiate into several mesenchymal lineages. The aim of this work was to investigate the possibility of isolating pluripotent precursor cells from equine peripheral blood and compare them with equine bone marrow-derived mesenchymal stem cells. Human mesenchymal stem cells (MSCs) were used as a control for cell multipotency assessment. Venous blood (n ‫؍‬ 33) and bone marrow (n ‫؍‬ 5) were obtained from adult horses. Mononuclear cells were obtained by Ficoll gradient centrifugation and cultured in monolayer, and adherent fibroblast-like cells were tested for their differentiation potential. Chondrogenic differentiation was performed in serum-free medium in pellet cultures as a threedimensional model, whereas osteogenic and adipogenic differentiation were induced in monolayer culture. Evidence for differentiation was made via biochemical, histological, and reverse transcription-polymerase chain reaction evaluations. Fibroblast-like cells were observed on day 10 in 12 out of 33 samples and were allowed to proliferate until confluence. Equine peripheral blood-derived cells had osteogenic and adipogenic differentiation capacities comparable to cells derived from bone marrow. Both cell types showed a limited capacity to produce lipid droplets compared to human MSCs. This result may be due to the assay conditions, which are established for human MSCs from bone marrow and may not be optimal for equine progenitor cells. Bone marrow-derived equine and human MSCs could be induced to develop cartilage, whereas equine peripheral blood progenitors did not show any capacity to produce cartilage at the histological level. In conclusion, equine peripheral blood-derived fibroblast-like cells can differentiate into distinct mesenchymal lineages but have less multipotency than bone marrow-derived MSCs under the conditions used in this study.

In Vitro Studies of Horse Umbilical Cord Matrix-Derived Cells: From Characterization to Labeling for Magnetic Resonance Imaging

“…Committed equine UCM-derived populations gave rise to both chondrogenic and adipogenic lineages, although the literature suggests that equine MSCs show inefficient differentiation into adipogenic lineages compared to human and rodent MSCs. This discrepancy may be due to the heterogeneity of the starting population [30] and/ or culture conditions [23] in previous studies. Furthermore, in this study we were able to show morphologically neuron-like cells and expression of specific neuronal markers after differentiation in vitro.…”

Mesenchymal stem cells from amnion and amniotic fluid in the bovine

“…As in humans, also in veterinary medicine, mesenchymal stem cells (MSCs) have been harvested successfully from a wide range of tissues (Smith et al 2003, Kern et al 2006, Koerner et al 2006, Arnhold et al 2007, Giovannini et al 2008, Vidal et al 2008. The most characterized sources of MSCs are bone marrow (Fortier et al 1998, Ringe et al 2002, Mitchell et al 2003, Smith et al 2003, Kern et al 2006, Vidal et al 2006, Arnhold et al 2007, Vidal et al 2008) and the adipose tissue (Kern et al 2006, Vidal et al 2008; however, the procedures employed to isolate these tissues are still invasive and cells are usually obtained with low efficiency.…”

Coleta de células progenitoras periféricas por aférese automatizada em equino: relato de procedimento

“…Atualmente, as técnicas utilizadas para obtenção de células-tronco em equinos é a coleta de células mesequimais, principalmente provenientes de tecido adiposo e medula óssea. Alguns autores afirmam que a medula óssea é a fonte mais rica para produção de células pluripotentes e utilização em terapias reconstrutivas de equinos e outras espécies (Koerner et al, 2006;Hegewald et al, 2004). No entanto, a coleta de CPP pode apresentar-se como método menos invasivo para obtenção dessas células, o que implicaria menor desconforto para o paciente e maior facilidade para aplicação clínica.…”

“…A coleta de CPP em humanos já é uma realidade em centros de hemoterapia, e na Europa já substituem os transplantes de medula óssea convencionais (Corgozinho et al, 2012). Ainda, estudos de células-tronco isoladas de sangue periférico em cães e cobaias têm demonstrado grande capacidade de diferenciação em células mesenquimais para uso terapêutico (Koerner et al, 2006).…”

“…As vantagens da coleta de CPP por aférese são a possibilidade da realização da coleta em regime ambulatorial, não sendo necessária a anestesia do paciente; a menor taxa de morbidade e mortalidade; a menor contaminação de células tumorais no produto, além de representar um método menos invasivo e mais seguro quando comparado à coleta de células-tronco derivadas da medula óssea ou de tecido adiposo (Stem..., 2013;Koerner et al, 2006).…”