J Med Assoc Thai 2010; 93 (12):183

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Isolation, Characterization and Neural Differentiation Potential of Amnion Derived Mesenchymal Stem Cells
Manochantr S Mail, Tantrawatpan C , Kheolamai P , U-pratya Y , Supokawej A , Issaragrisil S

Mesenchymal stem cells (MSCs) derived from amnion are considered to be adult stem cells that can be easily
obtained in large quantities by a less invasive method in comparison to bone marrow-derived MSCs (BM-MSCs). However,
the biological properties and the differentiation capacity of amnion-derived MSCs (AM-MSCs) are still poorly characterized.
The objectives of this study were to isolate, characterize and explore the potential of AM-MSCs in differentiating toward neural
lineage in comparison to those of BM-MSCs. To isolate AM-MSCs, amnion was digested with trypsin-EDTA and cultured in
Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum. The expression profiles of several
MSC markers were examined by flow cytometry. AM-MSCs from passage 3-5 were used for adipogenic, osteogenic and
neural differentiation assays by culturing in appropriate induction media. The expression of several neural marker genes,
including MAP-2, GFAP and β-tubulin III in AM-MSCs was determined by quantitative real time-PCR. The expression of
neural-specific markers, MAP-2 and β-tubulin III, was subsequently confirmed by immunocytochemistry using confocal laser
microscope. The results demonstrated that AM-MSCs could be easily expanded to 18-20 passages while maintaining the
undifferentiated state and exhibiting MSC markers (CD73, CD90, and CD105) but do not express the hematopoietic markers
(CD34 and CD45). Similar to BM-MSCs, AM-MSCs were able to differentiate to several mesodermal-lineages including
adipocytes and osteoblasts. Moreover, these cells could be induced to differentiate to neuron-like cells as characterized by cell
morphology and the expression of several neural markers including MAP-2, GFAP and β-tubulin III. The present study
demonstrated that AM-MSCs can be easily obtained and expanded in culture. These cells also have transdifferentiation
capacity as evidenced by their neural differentiation potential. According to the results, amnion can be used as an alternative
source of MSCs for stem cell therapy in neurodegenerative diseases.

Keywords: Mesenchymal stem cell, Amnion, Neural differentiation

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