Hanging drop culture enhances differentiation of human adipose‐derived stem cells into anterior neuroectodermal cells using small molecules

International Journal of Developmental Neuroscience - Tập 59 - Trang 21-30 - 2017
Noushin Amirpour1, Shahnaz Razavi1, Ebrahim Esfandiari1, Batoul Hashemibeni1, Mohammad Kazemi2, Hossein Salehi1
1Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
2Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Tóm tắt

AbstractInspired by in vivo developmental process, several studies were conducted to design a protocol for differentiating of mesenchymal stem cells into neural cells in vitro. Human adipose‐derived stem cells (hADSCs) as mesenchymal stem cells are a promising source for this purpose. At current study, we applied a defined neural induction medium by using small molecules for direct differentiation of hADSCs into anterior neuroectodermal cells. Anterior neuroectodermal differentiation of hADSCs was performed by hanging drop and monolayer protocols. At these methods, three small molecules were used to suppress the BMP, Nodal, and Wnt signaling pathways in order to obtain anterior neuroectodermal (eye field) cells from hADSCs. After two and three weeks of induction, the differentiated cells with neural morphology expressed anterior neuroectodermal markers such as OTX2, SIX3, β‐TUB III and PAX6. The protein expression of such markers was confirmed by real time, RT‐PCR and immunocytochemistry methods According to our data, it seems that the hanging drop method is a proper approach for neuroectodermal induction of hADSCs. Considering wide availability and immunosuppressive properties of hADSCs, these cells may open a way for autologous cell therapy of neurodegenerative disorders.

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International Journal of Developmental Neuroscience

Tập 59

21-30

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