Method for preparing animal mature neuron single cells

Abstract

The invention discloses a method for preparing animal mature neuron single cells, and belongs to the technical field of separation of mature neuron single cells. According to the method, firstly, D-PBS is utilized for conducting perfusion on a mouse, blood cells are removed, and the influence of the blood cells on subsequent separation is avoided; Hibernate-A is utilized as a culture medium in thedigestion process, without considering the problem of early oxygen introduction or oxygen introduction in the digestion process, the time can be effectively saved by at least 30 minutes, and the damage to neurons is reduced; by means of the method, the single cells can be prepared without special instruments; through the method, oligodendrocytes, astrocytes and neurons can be simultaneously separated out, sequencing or function analysis can be conducted on different types of nerve cells in the same batch of samples, the experimental errors can be reduced, and the cost is reduced.

Description

Technical field [0001] The invention relates to a method for preparing animal mature neuron single cells, and belongs to the technical field of mature neuron single cell separation. Background technique [0002] In the organism, every cell is unique, even the same type of cells in the same organ or tissue are all heterogeneous. As the most important organ of the organism, the brain has its own unique characteristics. For example, mature neurons are highly differentiated and cannot undergo cell division. This may lead to many neurodegenerative and mental diseases, such as Alzheimer’s disease , Parkinson's syndrome, etc. [0003] Through single-cell sequencing, it is possible to understand the heterogeneity between cells, find specific groups of neurons that are specifically involved in a certain disease, and at the same time reclassify neurons in a certain brain area. It turns out that due to technical limitations, neurons Relatively speaking, the classification is not perfect, si...

AI Technical Summary

Problems solved by technology

At present, there is only one kit for isolating mature neurons in the market, that is, the kit for isolating mature neurons from rats and mice produced by Miltenyi. At 300,000-500,000 / unit, at the same time, the final separation of blood cells in this kit will reduce the yield of nerve cells, so its usage rate is very low, and there are few articles published using this kit.

Although there are many articles on the preparation of mature neurons that can be referred to, there are problems such as the need for special equipment, the inability to completely digest and separate nerve cells, and the inability to separate neurons and different types of nerve cells at the same time.

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