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KMID : 0894520080120010031
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Development & Reproduction
2008 Volume.12 No. 1 p.31 ~ p.39
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Differentiation of Dopaminergic and Cholinergic Neurons from Mesenchymal-like Stem Cells Derived from the Adipose Tissue
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Hong In-Kyung
Jeong Na-Hee
Kim Ju-Ran
Do Byung-Rok
Kim Hae-kwon
Kang Sung-Goo
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Abstract
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Neural tissue has limited intrinsic capacity of repair after injury, and the identification of alternate sources of neural stem cells has broad clinical potential. We isolated mesechymal-like stem cells from human adipose tissues (AT-MSCs), and studied on transdifferentiation-promoting conditions in neural cells. Dopaminergic and cholinergic neuron induction of AT-MSCs was also studied. Neural differentiation was induced by adding bFGF, EGF, dimethyl sulphoxide (DMSO) and butylated hydroxyanisole (BHA) in N2 Medium and N2 supplement The immunoreactive cells for β-tubulin Ⅲ, a neuron-specific marker, GFAP, an astrocyte marker, or Gal-C, an oligodendrocyte marker, were found. AT-MSCs treated with bFGF, SHH and FGF8 were differentiatied into dopaminergic neurons that were immunopositive for TH antibody. Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor (bFGF), retinoic acid (RA) and sonic hedgehog (Shh). AT-MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including neuro Dl, β-tubulin Ⅲ, GFAP and nestinwas markedly elevated during this acute differentiation. While the stem cell markers such as SCF, C-kit, and Stat-3 were not expressed after preinduction medium culture, we confirmed the differentiation of dopaminergic and cholinergic neurons by TH/ β-tubulin Ⅲ or ChAT/ β-tubulin Ⅲ positive cells. Conclusively, AT-MSCs can be differentiated into dopaminergic and cholinergic neuronsand these findings suggest that AT-MSCs are alternative cell source of treatment for neurodegenerative diseases.
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KEYWORD
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Human adipose tissue, Mesenchymal stem cells, Cholinergic neuron, Dopaminergic neuron
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