Method for inducing differentiation of adipose-derived stem cells into dopaminergic neurons

Abstract

The invention discloses a method for inducing differentiation of adipose-derived stem cells into dopaminergic neurons, and belongs to the technical field of biomedicine. The dopaminergic neurons are obtained through separation of the adipose-derived stem cells, subculture of the adipose-derived stem cells and induced culture and conversion, and a large number of the adipose-derived stem cells areobtained through separation and culture of adipose tissue from autologous sources; the adipose-derived stem cells are converted into the dopaminergic neurons through two specific culture media, batches of stable dopaminergic neurons can be obtained safely and effectively, and a high differentiation rate is achieved. It is shown through immunofluorescence and cell counting experiments that the conversion rate of the dopaminergic neurons is 90.4+ / -0.76, the survival rate is 98.9+ / -0.55, and the average amount of dopamine which is secreted by the converted dopaminergic neurons and is detected byan ELISA kit is 450 pg / ml; and therefore, it is confirmed that the steps of extraction and conversion of the adipose-derived stem cells are gentle and stable, and the extracted cells are suitable formass application.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to a method for inducing fat stem cells to differentiate into dopaminergic neurons. Background technique [0002] Parkinson's disease (PD) is a common neurodegenerative disease characterized by the disappearance of dopaminergic neurons in the substantia nigra and the accumulation of Lewy bodies, muscle rigidity and slow movement. , dyskinesia, resting tremor, and abnormal sleep behavior are clinical manifestations. In my country, the prevalence of Parkinson's disease among middle-aged and elderly people over 65 years old is 1.7%. With the aging phenomenon increasing year by year, the diagnosis and treatment of Parkinson's patients should attract great attention. At present, the main treatment plan for Parkinson's is to promote the release of dopamine and relieve motor function. Drug therapy can relieve symptoms to a certain extent, but long-term use produces drug tolerance, and...

AI Technical Summary

Problems solved by technology

One of the clinical treatment options is drug therapy. Patients can relieve clinical manifestations by taking dopamine substitutes. However, this method has limited curative effect. large, causing the human body to be unable to tolerate it, and the treatment of the patient's disease is ineffective; another treatment method is deep brain stimulation (DBS), commonly known as a brain pacemaker. DBS stimulation schemes are different due to individual differences. When choosing this treatment method, there are strict indications for patient selection, and the debugging of the controller is more complicated, which may easily lead to overreaction of the cell switch and side effects, and the fixed parameters cannot adapt to the patient's condition when the condition changes. The need to change, sometimes the treatment effect is limited, and the effect of DBS application gradually weakens or disappears after a period of time, and at the same time, no effective treatment can be obtained

At present, researches at home and abroad use fetal brain neural stem cells, embryonic neural stem cells, iPSCs cells, etc. for transplantation to treat Parkinson's disease. Cells are tumorigenic, and this method has problems such as complicated technology, high cost, difficult control of experiments, and uneven purity between batches. There are many public opinions, which limit the application of the above stem cell method and make it difficult to further develop