Establishment And Optimization Of A New Neural Differentiation System Derived From Human ES/iPs Cells

Due to the disease,age and trauma,it often leads to death and pathological changes of neurons.However,the traditional idea is that the neurons in the human body can not be regenerated,which leads to neurological diseases,it has long puzzled clinical medicine.The occurrence of nervous system diseases usually causes patients to have obstacles in consciousness,movement and sensation,which often cause great distress to patients' lives.With the continuous development of biology and medical research,the advent of stem cell therapy has brought hope for the cure of nervous system diseases.We hope that neural stem cell transplantation can be used to treat nervous system diseases.Because of the inability to directly obtain neural stem cells from the human body,we can only use cells to induce differentiation in vitro to obtain neural stem cells.Through the continuous study of the development of the nervous system in the body by the animal model and the exploration of the system of neural differentiation in vitro,it has been able to successfully obtain neural stem cells through in vitro induction,including a variety of neurosegregation systems with ES/i PS cells as the starting cells and the differentiation of various somatic cells.A system of neural transdifferentiation of the beginning cells.In this study,human ES and i PS cells were used as differentiation initiating cells,and neural precursor cells(NPCs)were induced in vitro.According to the previous research basis,this study has created a new induction method through the replacement of the cell culture matrix in the process of cell culture and the optimization of the differentiation medium.The experiment was divided into two groups: 1.ES cells as the differentiation initiation cells,and 2.i PS cells as the differentiation initiation cells.At the cell level in vitro,using real-time quantitative PCR,flow cytometry,immunofluorescence and other methods to analyze the downstream differentiation ability of the cells at different time points in the differentiation process,and to evaluate the electrical signal conduction ability of the neuron cells by the patch clamp technique.In the experiment,the new SD rat structure was used in the experiment.A lateral ventricle transplantation model was established to detect the viability,differentiation and migration of cells in vivo.The experimental results showed that: first,the structure of the nerve rosette appeared in the differentiation process of UC1-i PS cells to NPCs,and the results of flow cytometry showed that the positive rate of Pax6 was about 80%,the positive rate of Nestin was about 90%,the NPCs was differentiated efficiently,the differentiation process lasted 7 days and the differentiation was rapid,and the H1-ES/UC1-i PS cells were differentiated into NPCs.Through the detection of H1-ES/UC1-i PS cells and the samples of D1,D3,D5 and D7 cells during differentiation,the expression of Nanog in the pluripotent gene decreased with the process of differentiation,and the high expression of Sox2 due to its high expression in ES/i PS cells and NSCs all the time in the differentiation system,and Nestin,Sox1,Pax6,etc.The specific gene expression is up-regulated;secondly,the NPCs induced by the new neural differentiation system can well differentiate into different kinds of neuron cells and glia cells in vitro,and third,the deity cells of the NPCs downstream differentiation induced by the new neural differentiation system have good electrical signal conduction energy.Fourth,through the experiment of lateral ventricle transplantation in newborn rats,it was found that the NPCs induced by new neural differentiation system could survive,differentiate and migrate in animals.In conclusion,we have successfully established a new neural differentiation system,which is stable,fast and efficient.From the point of view of clinical application,the differentiation system has great application significance in the treatment of nervous system diseases.