Method for constructing three-dimensional engineered nerve tissue, three-dimensional engineered nerve tissue and application thereof

Abstract

The invention relates to a method for constructing three-dimensional engineered nervous tissue, the three-dimensional engineered nervous tissue and an application thereof. Process. The method comprises the following steps: 1, carrying out isolated culture on animal neural stem cells; 2, applying a physiological electric field; subculturing the neural stem cells isolated and cultured in the step 1to the fifth generation, centrifuging neural spheres with a diameter of 100-200 microns for later use, pre-melting matrix gel Matrigel on ice for later use, and continuously applying a physiological electric field to stimulate for 7-14 days through physiological electric field application equipment for 1-2 hours every day; 3, optimizing a culture solution suitable for engineering nerve tissue growth: adding 1 / 50 of B27, 1 / 100 of N2, 10-20 ng / ml of bFGF and 1% of fetal calf serum into DMEM / F12; and 4, applying the physiological electric field for 7-14 days to obtain the three-dimensional engineered nervous tissue. The technical problems that in the application process of neural tissue engineering, the number of neuronal differentiation of existing neural stem cells is extremely small, and neurite cannot be induced to grow orderly in a three-dimensional growth environment to form a developed neural network are solved.

Description

technical field [0001] The invention relates to a method for constructing a three-dimensional engineered nerve tissue, a three-dimensional engineered nerve tissue and uses thereof. Background technique [0002] There are about 133,000 to 262,000 new spinal cord injury patients in the world every year. The economic cost of nursing and treatment due to illness and paralysis is as high as tens of billions of dollars, which has caused a huge burden on social development. Nervous system injury leads to tissue disintegration and cell apoptosis at the injured site, and the functions carried by the corresponding structures are also damaged or lost. Engineered neural tissue is a multidisciplinary interdisciplinary field emerging in recent years, integrating biology, materials science, informatics and clinical medicine. Engineered nerve tissue is an ordered nerve tissue with a three-dimensional tissue structure constructed in vitro with nerve cells as seed cells and bioactive materia...

AI Technical Summary

Problems solved by technology

[0005] The present invention provides a method for constructing three-dimensional engineered neural tissue to solve the problem of the extremely small number of neuron differentiation existing in the application process of neural stem cells in the process of neural tissue engineering and the inability to induce the orderly growth and formation of neurites in a three-dimensional growth environment Technical Issues of Developed Neural Networks

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