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Isolation and characterization of equine amnion mesenchymal stem cells.

Coli A, Nocchi F, Lamanna R, Iorio M, Lapi S, Urciuoli P, Scatena F, Giannessi E, Stornelli MR, Passeri S - Cell Biol Int Rep (2010) (2011)

Bottom Line: The cells were grown in α-MEM (α-modified minimum essential medium) and the effect of EGF (epidermal growth factor) supplementation was evaluated.EGF supplementation of the culture medium resulted in a significant increase in PD growth parameter and in the formation of bone nodules for the osteogenic differentiation.By immunohistochemistry the amnion tissue shows a positivity for the c-Kit (cluster tyrosine-protein kinase), CD105 and Oct-4 (octamer-binding transcription factor 4) antigens that confirmed the presence of MSCs with embryonic phenotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Pathology, University of Pisa, Pisa, Italy.

ABSTRACT
The amnion is a particular tissue whose cells show features of multipotent stem cells proposed for use in cellular therapy and regenerative medicine. From equine amnion collected after the foal birth we have isolated MSCs (mesenchymal stem cells), namely EAMSCs (equine amnion mesenchymal stem cells), from the mesoblastic layer. The cells were grown in α-MEM (α-modified minimum essential medium) and the effect of EGF (epidermal growth factor) supplementation was evaluated. To assess the growth kinetic of EAMSCs we have taken into account some parameters [PD (population doubling), fold increase and DT (doubling time)]. The differentiation in chondrogenic, adipogenic and osteogenic types of cells and their epitope expression by a cytofluorimetric study have been reported. EGF supplementation of the culture medium resulted in a significant increase in PD growth parameter and in the formation of bone nodules for the osteogenic differentiation. By immunohistochemistry the amnion tissue shows a positivity for the c-Kit (cluster tyrosine-protein kinase), CD105 and Oct-4 (octamer-binding transcription factor 4) antigens that confirmed the presence of MSCs with embryonic phenotype.

No MeSH data available.


EAMSCs at the beginning of culture (A) and at confluence (B)
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Figure 1: EAMSCs at the beginning of culture (A) and at confluence (B)

Mentions: On average 1.2±0.6×107 (n = 4) nucleated cells were extracted from amnion and 8±3×105 (n = 4) MSCs were isolated at P0. In the first step of the culture, two types of cells were isolated: a spindle-shaped type and a round shape type which disappeared in the early passages. The cells reached confluence after 8 days in P0 (Figure 1). Mesenchymal cells cultured with α-MEM 10-10 reached senescence after ten passages with a CPD of 27.96±8.7.


Isolation and characterization of equine amnion mesenchymal stem cells.

Coli A, Nocchi F, Lamanna R, Iorio M, Lapi S, Urciuoli P, Scatena F, Giannessi E, Stornelli MR, Passeri S - Cell Biol Int Rep (2010) (2011)

EAMSCs at the beginning of culture (A) and at confluence (B)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475441&req=5

Figure 1: EAMSCs at the beginning of culture (A) and at confluence (B)
Mentions: On average 1.2±0.6×107 (n = 4) nucleated cells were extracted from amnion and 8±3×105 (n = 4) MSCs were isolated at P0. In the first step of the culture, two types of cells were isolated: a spindle-shaped type and a round shape type which disappeared in the early passages. The cells reached confluence after 8 days in P0 (Figure 1). Mesenchymal cells cultured with α-MEM 10-10 reached senescence after ten passages with a CPD of 27.96±8.7.

Bottom Line: The cells were grown in α-MEM (α-modified minimum essential medium) and the effect of EGF (epidermal growth factor) supplementation was evaluated.EGF supplementation of the culture medium resulted in a significant increase in PD growth parameter and in the formation of bone nodules for the osteogenic differentiation.By immunohistochemistry the amnion tissue shows a positivity for the c-Kit (cluster tyrosine-protein kinase), CD105 and Oct-4 (octamer-binding transcription factor 4) antigens that confirmed the presence of MSCs with embryonic phenotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Pathology, University of Pisa, Pisa, Italy.

ABSTRACT
The amnion is a particular tissue whose cells show features of multipotent stem cells proposed for use in cellular therapy and regenerative medicine. From equine amnion collected after the foal birth we have isolated MSCs (mesenchymal stem cells), namely EAMSCs (equine amnion mesenchymal stem cells), from the mesoblastic layer. The cells were grown in α-MEM (α-modified minimum essential medium) and the effect of EGF (epidermal growth factor) supplementation was evaluated. To assess the growth kinetic of EAMSCs we have taken into account some parameters [PD (population doubling), fold increase and DT (doubling time)]. The differentiation in chondrogenic, adipogenic and osteogenic types of cells and their epitope expression by a cytofluorimetric study have been reported. EGF supplementation of the culture medium resulted in a significant increase in PD growth parameter and in the formation of bone nodules for the osteogenic differentiation. By immunohistochemistry the amnion tissue shows a positivity for the c-Kit (cluster tyrosine-protein kinase), CD105 and Oct-4 (octamer-binding transcription factor 4) antigens that confirmed the presence of MSCs with embryonic phenotype.

No MeSH data available.