Method for inducing human spinal cord motor nerve precursor cells to differentiate into spinal cord motor neurons

Abstract

The invention discloses a method for inducing human spinal cord motor nerve precursor cells to differentiate into spinal cord motor neurons. The method comprises the following steps: 1) culturing of spinal cord motor nerve precursor cells: the spinal cord motor nerve precursor cells are disassociated into single cells, a coated culture dish is inoculated with the single cells for adherent culture,wherein a culture solution is a spinal cord motor nerve precursor cell culture medium, and the single cells are cultured in an incubator at the temperature of 37 DEG C and 5% CO2 saturated humidity;2) induction of the spinal cord motor neurons: the spinal cord motor neuron precursor cells obtained in the step 1) are cultured in a basal complete culture medium, different space-time specific signal channels are added at different time to control induced differentiation of small molecules and / or growth factors, the cells are cultured in the incubator at the temperature of 37 DEG C and 5% CO2 saturated humidity for 3-30 days, and the solution is changed once every two days. The method is efficient, fast, stable and safe, and 70% or more spinal cord motor neurons can be obtained on the thirdday of differentiation.

Description

technical field [0001] The invention relates to the technical field of cell differentiation and culture. More specifically, it relates to a method for inducing the differentiation of human spinal cord motor neuron precursor cells into spinal cord motor neurons. Background technique [0002] In recent years, the number of spinal cord injury patients due to car accidents, falls, contusions and other reasons has been increasing year by year. Due to damage to the internal nerves of the spinal cord, patients with spinal cord injury generally suffer from paralysis of the lower limbs, making it difficult to take care of themselves. Under normal physiological conditions, damaged nerve cells in the human body cannot repair themselves, so traditional medical methods are difficult to completely cure the disease. Cell replacement therapy provides a new therapeutic approach for patients with spinal cord injury. Spinal motoneurons are one of the ideal cell types for disease treatment o...

AI Technical Summary

Problems solved by technology

The traditional differentiation of neural precursor cells into neurons generally has the problems of long differentiation time and low differentiation efficiency

Moreover, the use of animal-derived components and extracted proteins will be involved in the differentiation process. Animal-derived components will potentially introduce the risk of pathogenic microorganisms. It is difficult to control the uniformity of extracted protein batches, which will lead to instability of the differentiation system.

In the traditional process of differentiation of spinal cord motor nerve precursor cells into spinal cord motor neurons, the above problems also exist. The differentiation time needs at least seven days, and the differentiation efficiency is less than 40%.

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