Isolation and Characterization of Canine Amniotic Membrane-Derived Multipotent Stem Cells

Abstract

Recent studies have shown that amniotic membrane tissue is a rich source of stem cells in humans. In clinical applications, the amniotic membrane tissue had therapeutic effects on wound healing and corneal surface reconstruction. Here, we successfully isolated and identified multipotent stem cells (MSCs) from canine amniotic membrane tissue. We cultured the canine amniotic membrane-derived multipotent stem cells (cAM-MSCs) in low glucose DMEM medium. cAM-MSCs have a fibroblast-like shape and adhere to tissue culture plastic. We characterized the immunophenotype of cAM-MSCs by flow cytometry and measured cell proliferation by the cumulative population doubling level (CPDL). We performed differentiation studies for the detection of trilineage multipotent ability, under the appropriate culture conditions. Taken together, our results show that cAM-MSCs could be a rich source of stem cells in dogs. Furthermore, cAM-MSCs may be useful as a cell therapy application for veterinary regenerative medicine.

Differential developmental competence and gene expression patterns in buffalo (Bubalus bubalis) nuclear transfer embryos reconstructed with fetal fibroblasts and amnion mesenchymal stem cells

“…It is a well-known fact that mixed population of cells cultured from amniotic membrane in many species such as human, ovine, chicken, rat, equine and canine also displayed heterogeneous cell population (Marcus et al 2008;Mauro et al 2010;Bacenkova et al 2011;Coli et al 2011;Gao et al 2012;Park et al 2012). In agreement with these reports, it was observed that cultured cells exhibited both spindleshaped fibroblasts and polygonal shaped epithelial cell-like morphology in vitro.…”

“…These results indicate that these cells were in an undifferentiated state. Expression of these pluripotency marker genes has previously been reported in amnion-derived cells of various mammalian species including rat (Marcus et al 2008), canine (Park et al 2012) and chicken (Gao et al 2012). In addition, reverse transcriptase PCR results indicated that AMSCs expressed the mesenchymal stem cells markers CD44 and CD29, but not haematopoietic surface markers such as CD34 as reported earlier in bovine .…”

“…Recently, Yamahara et al (2014) reported that mesenchymal stem cells isolated from human amniotic tissue have an angiogenesis potential. In animals, amnion-derived stem cells have potential of differentiation into multilineage mesenchymal cell types in many species such as bovine , canine (Park et al 2012), feline (Rutigliano et al 2013), chicken (Gao et al 2012), rat (Marcus et al 2008) and equine (Coli et al 2011;Violini et al 2012;Seo et al 2013). Further, the amnion derived MSCs are reported to express certain key pluripotency markers including OCT4 .…”

Membrana amniótica: fonte alternativa de células-tronco mesenquimais em diversas espécies animais

“…Células-tronco multipotentes foram isoladas e identificadas em membranas amnióticas de cães, caracterizando o âmnio como uma rica fonte de células-tronco, com possíveis utilizações em terapia celular na medicina veterinária regenerativa (Park et al 2012). MSCs coletadas do fluído amniótico de fetos caninos no terceiro trimestre gestacional apresentaram diferenciação in vitro em linhagens celulares condrogênicas, adipogênicas, osteogênicas e neurogênicas, demonstrando potencial progenitor (Fernandes et al 2012).…”

Comparação molecular entre células-tronco mesenquimais de membrana amniótica de cão e de gato

“…Muitas pesquisas estão em andamento com a intenção de explorar estas características visando finalidades terapêuticas (BYDLOWSKI et al, 2009a;PARK et al, 2012;VIDANE et al, 2013). As CTs podem ser totipotentes, pluripotentes ou multipotentes.…”