TY - JOUR
T1 - Characterization and potential applications of progenitor-like cells isolated from horse amniotic membrane
AU - Lange-Consiglio, A.
AU - Corradetti, B.
AU - Bizzaro, D.
AU - Magatti, M.
AU - Ressel, L.
AU - Tassan, S.
AU - Parolini, Ornella
AU - Cremonesi, F.
PY - 2012
Y1 - 2012
N2 - The aim of this work was to isolate, for the first time, progenitor-like cells from the epithelial (AECs) and mesenchymal (AMCs) portions of the horse amniotic membrane, and to define the biological properties of these cells. AECs displayed polygonal epithelial morphology, while AMCs were fibroblast-like. Usually, six to eight passages were reached before proliferation decreased, with 13.08 and 26.5 cell population doublings attained after 31 days for AECs and AMCs, respectively. Immunocytochemical studies performed at passage 3 (P3) showed that both cell populations were positive for the expression of specific embryonic markers (TRA-1-60, SSEA-3, SSEA-4 and Oct-4). Meanwhile, RT-PCR performed at P1 and P5 showed expression of mesenchymal stem/stromal cell markers (CD29, CD105, CD44 and CD166) with negativity for CD34 at P1, although this marker began to be expressed by P5. The cells also expressed MHC-I at both P1 and P5, but lacked MHC-II expression at P1. Both AECs and AMCs demonstrated high plasticity, differentiating in vitro toward the osteogenic, adipogenic, chondrogenic and neurogenic lineages. Equine amnion-\derived cells could also be frozen and recovered without loss of their functional integrity in terms of morphology, presence of specific stemness markers and differentiation ability, although the renewal capacity was lower than that observed for freshly isolated cells. To investigate potential therapeutic effects and cell tolerance in vivo, horse amnion-derived cells were allogeneically injected into three horses with tendon injuries, resulting in a quick reduction in tendon size and ultrasonographic cross-sectional area measurements. These results suggest that horse amnion-derived cells may be useful for cell therapy applications.
AB - The aim of this work was to isolate, for the first time, progenitor-like cells from the epithelial (AECs) and mesenchymal (AMCs) portions of the horse amniotic membrane, and to define the biological properties of these cells. AECs displayed polygonal epithelial morphology, while AMCs were fibroblast-like. Usually, six to eight passages were reached before proliferation decreased, with 13.08 and 26.5 cell population doublings attained after 31 days for AECs and AMCs, respectively. Immunocytochemical studies performed at passage 3 (P3) showed that both cell populations were positive for the expression of specific embryonic markers (TRA-1-60, SSEA-3, SSEA-4 and Oct-4). Meanwhile, RT-PCR performed at P1 and P5 showed expression of mesenchymal stem/stromal cell markers (CD29, CD105, CD44 and CD166) with negativity for CD34 at P1, although this marker began to be expressed by P5. The cells also expressed MHC-I at both P1 and P5, but lacked MHC-II expression at P1. Both AECs and AMCs demonstrated high plasticity, differentiating in vitro toward the osteogenic, adipogenic, chondrogenic and neurogenic lineages. Equine amnion-\derived cells could also be frozen and recovered without loss of their functional integrity in terms of morphology, presence of specific stemness markers and differentiation ability, although the renewal capacity was lower than that observed for freshly isolated cells. To investigate potential therapeutic effects and cell tolerance in vivo, horse amnion-derived cells were allogeneically injected into three horses with tendon injuries, resulting in a quick reduction in tendon size and ultrasonographic cross-sectional area measurements. These results suggest that horse amnion-derived cells may be useful for cell therapy applications.
KW - Amnion
KW - Animals
KW - Biological Assay
KW - Cell Differentiation
KW - Cell Proliferation
KW - Cell Separation
KW - Cell Shape
KW - Colony-Forming Units Assay
KW - Epithelial Cells
KW - Female
KW - Horses
KW - Immunohistochemistry
KW - Mesenchymal Stromal Cells
KW - Multipotent Stem Cells
KW - Reverse Transcriptase Polymerase Chain Reaction
KW - Rupture
KW - Staining and Labeling
KW - Stem Cell Transplantation
KW - Stem Cells
KW - Tendons
KW - Amnion
KW - Animals
KW - Biological Assay
KW - Cell Differentiation
KW - Cell Proliferation
KW - Cell Separation
KW - Cell Shape
KW - Colony-Forming Units Assay
KW - Epithelial Cells
KW - Female
KW - Horses
KW - Immunohistochemistry
KW - Mesenchymal Stromal Cells
KW - Multipotent Stem Cells
KW - Reverse Transcriptase Polymerase Chain Reaction
KW - Rupture
KW - Staining and Labeling
KW - Stem Cell Transplantation
KW - Stem Cells
KW - Tendons
UR - http://hdl.handle.net/10807/92413
U2 - 10.1002/term.465
DO - 10.1002/term.465
M3 - Article
SN - 1932-6254
VL - 6
SP - 622
EP - 635
JO - Journal of Tissue Engineering and Regenerative Medicine
JF - Journal of Tissue Engineering and Regenerative Medicine
ER -