Mesenchymal stem cells from amnion and amniotic fluid in the bovine

Abstract

Amnion and amniotic fluid (AF) are noncontroversial and inexhaustible sources of mesenchymal stem cells (MSCs) that can be harvested noninvasively at low cost. As in humans, also in veterinary field, presumptive stem cells derived from these tissues reveal as promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. The aim of this work is to obtain and characterize, for the first time in bovine species, presumptive MSCs from the epithelial portion of the amnion (AECs) and from the AF (AF-MSCs) to be used for clinical applications. AECs display a polygonal morphology, whereas AF-MSCs exhibit a fibroblastic-like morphology only starting from the second passage, being heterogeneous during the primary culture. For both lines, the proliferative ability has been found constant over the ten passages studied and AECs show a statistically lower (P!0.05) doubling time with respect to AF-MSCs. AECs express MSC-specific markers (ITGB1 (CD29), CD44, ALCAM (CD166), ENG (CD105), and NT5E (CD73)) from P1 to P3; in AF-MSCs, only ITGB1, CD44, and ALCAM mRNAs are detected; NT5E is expressed from P2 and ENG has not been found at any passage. AF-MSCs and AECs are positive for the pluripotent markers (POU5F1 (OCT4) and MYC (c-Myc)) and lack of the hematopoietic markers. When appropriately induced, both cell lines are capable of differentiating into ectodermal and mesodermal lineages. This study contributes to reinforce the emerging importance of these cells as ideal tools in veterinary medicine. A deeper evaluation of the immunological properties needs to be performed in order to better understand their role in cellular therapy.

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

“…Fibroblastlike appearing adherent MSCs isolated from buffalo amnion were successfully induced to multipotential differentiation into adipogenic, chondrogenic, and osteogenic lineages under specific culture conditions. The morphological features of MSCs were similar to the earlier observations of Corradetti et al (2013). It is widely accepted that multiple cell types can be derived by culturing AMSCs under appropriate conditions.…”

Enhanced osteogenic potential of mesenchymal stem cells from cortical bone: a comparative analysis

“…Colony-forming unit assay was performed on freshly isolated CBF-MSCs, AD-MSCs, and BM-MSCs as previously reported [12,17,48]. Briefly, cells at P2 were plated in six-well plates at different densities (100, 250, 500, and 1000 cells/cm 2 ) and cultured over a 15-day period.…”

“…MSCs from the bone marrow (BM-MSCs) were originally described by Friedenstein et al as non-phagocytic and non-hematopoietic cells and adherent to plastic [4]. Over the last 15 years, there has been an explosion in the reports of MSCs isolated from a variety of other adult sources, including skin [5], adipose tissue [6], umbilical cord blood [7,8] and matrix, peripheral blood [9], tendons [10,11], amnion [12][13][14], and bone [15][16][17]. Their plasticity, immunosuppressive potential, immuno-modulatory properties [18,19], and trophic activity [20,21] make MSCs critical players in tissue homeostasis.…”

Bone Morphogenetic Protein-12 Induces Tenogenic Differentiation of Mesenchymal Stem Cells Derived from Equine Amniotic Fluid

“…Some cells at P1 did not show AP staining. It might be due to the fact that during initial cultures, there is a heterogeneous population of cells in AF and in subsequent passages, nonproliferative cells are eliminated and MSCs form a uniform monolayer and thus show uniform AP expression [Arnhold et al, 2011;Corradetti et al, 2013].…”