Method for increasing yield of neural stem cells in adult nerve tissues by utilizing organotypic culture

Abstract

The invention relates to a method for increasing the yield of neural stem cells in adult nerve tissues by utilizing organotypic culture. The method is characterized in that the method comprises organotypic culture and cell culture, and all the operations are completed on an aseptic operating platform. The method has the advantages and positive effects that organotypic culture is adopted and then papain digestion and blowing and beating dispersion are carried out seven days later, so that the quantity of obtained living cells is higher than that of the living cells obtained through direct enzymatic digestion by more than three times; another participation factor is mass multiplication of the neural stem cells in the organotypic culture process, thus increasing the total of the stem cells; another technological improvement is that the cells are directly inoculated in a poly-L-ornithine (PLO) coated cell culture plate after tissue dispersion, and tissue residues (including cell debris) are removed several hours later by changing a culture solution, thus omitting the time-consuming and tedious step of gradient centrifugation and also avoiding the cytotoxicity caused by most gradient centrifugation solutions.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for increasing the harvest rate of neural stem cells in adult neural tissue by using organ culture. Background technique [0002] Neural stem cells (NSCs) are pluripotent stem cells that can differentiate into any kind of nerve cells. Neural precursor cells (NPC) are initially differentiated neural stem cells that can only differentiate into neurons. These two types of cells are generally referred to as neural stem cells, and this term is used in the present invention for the sake of brevity. [0003] The research results mainly derived from this century show that neural stem cells (neural stem cells, NSCs) and neural precursor cells (neural precursor cells, NPCs) not only exist in human embryonic development, but also exist in a small amount in some brain regions of adults and the center of the spinal cord. These neural stem cells can be extracted from donated cadavers f...

AI Technical Summary

Problems solved by technology

However, since adult nerve tissue contains a large amount of fiber-based connective tissue components and tight connections between cells, the tissue structure is much harder than embryonic tissue, which makes the enzyme digestion effect poor, and the tissue is not easy to disperse when blown mechanically, resulting in tissue damage. Incomplete dispersion and strong dispersion seriously damage the cells, so the amount of viable cells obtained is very small (low yield)

[0005] In the prior art, there is a technical problem of extremely low yield of living cells due to the hard adult nerve tissue and poor enzymatic digestion, so a new method is urgently needed to solve this technical problem

[0006] Through the search, no patent publications related to the patent application of the present invention have been found

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