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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۷۷-۷۷
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عنوان انگلیسی Ps-131: Direct Differentiation of Human Pluripotent Stem Cells into Functional Motor Neurons Using Small Molecules
چکیده انگلیسی مقاله Objective: Motor neuron differentiation from human pluripotent stem cells (hPSC) is a goal in regenerative medicine for cell therapy in diseases that loss these cells. Most protocols have low efficiency in generate functional motoneurons, therefor small molecules are new idea for dominance this challenge. Materials and Methods: Under this approach we tried to use an efficient, short time and cost effective protocol than reported protocols to generate functional motoneurons from hPSC. Therefor we used small molecules Dorsomorphin, A8301, XAV, Retinoic acid and Purmorphamine in this protocol. Results: Immunofleoroscent staining and flow cytometry analysis of hESC-derived neural ectoderm (NE) showed these structures are 92.68% NESTIN+, 64.40% PAX6+, 82.11%% SOX1+ in a chemically defined adherent culture. After replating of hESC-derived NE, the differentiated cells are Tuj1+, MAP2+, HB9+ and ISL1+.level of gene expression evaluated with real time RTPCR in different stages of differentiation protocol. Electrophysiological properties of differentiated cells recorded by whole cell Patch clamp technique. hESC_drived motor neurons displayed voltage gated delay rectifier K+ and Na+-Ca2+ inward currents. Conclusion: Our findings suggested that, hESC-derived neurons expressed special motor neuron proteins such as HB9 and ISL1 but electrophysiological properties showed that these cells were not mature because the resting membrane potential was more positive than mature motor neuron and Na+-Ca2+ inward currents were not sufficient for firing action potentials. It seems that the cells should be more developed for functional maturation.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/933
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