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Cell Journal، جلد ۱۹، شماره ۴، صفحات ۵۴۵-۵۵۱
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عنوان انگلیسی In Vitro Differentiation of Neural-Like Cells from Human Embryonic Stem Cells by A Combination of Dorsomorphin, XAV939, and A8301
چکیده انگلیسی مقاله Objective: Motor neuron differentiation from human embryonic stem cells (hESC) is a goal in regenerative medicine for cell therapy in diseases that damage motor neurons. Most protocols have decreased efficiency in generating functional motor neurons. Small molecules present a new approach for overcoming this challenge. Materials and methods: We studied an efficient, rapid, cost-effective protocol compared to previously reported protocols in an attempt to generate functional motor neurons from hESCs. The current protocol used the small molecules dorsomorphin, A8301, and XAV939. Results: Immunofluorescence staining and flow cytometry analysis of hESC-derived neural ectoderm (NE) indicated that they were positive for NESTIN (92.68%), PAX6 (64.40%), and SOX1 (82.11%) in a chemically defined adherent culture. The replated human embryonic stem cell (hESC)-derived NE differentiated cells were positive for Tuj1, MAP2, HB9 and ISL1. We evaluated the gene expression levels with real-time RT-PCR at different stages of the differentiation protocol. Electrophysiological properties of the differentiated cells were recorded by the whole-cell patch clamp technique. The hESC-derived motor neurons showed voltage gated delay rectifier K+ and Na+-Ca2+ inward currents. Conclusion: The findings suggested that hESC-derived neurons expressed the specific motor neuron proteins HB9 and ISL1. The electrophysiological properties of these cells were immature because the resting membrane potential (RMP) was more positive than the mature motor neuron and the Na+-Ca2+ inward currents were inadequate for firing action potentials. The cells should be more developed for functional maturation.
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نویسندگان مقاله | Zahra Valizadeh-Arshad


| Ebrahim Shahbazi


| Shiva Hashemizadeh


| Azadeh Moradmand


| Meysam Jangkhah


| Sahar Kiani


نشانی اینترنتی http://celljournal.org/journal/article/abstract/4232
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