A method for isolating nucleus pulposus primary cells

Abstract

The present disclosure relates to a method for efficiently separating nucleus pulposus primary cells, the method includes the following steps: a digestion step: the active components of the digestion solution used in this step include elastase, type II collagenase and glycosidase. Compared with the traditional separation method of primary nucleus pulposus cells, the method of the present disclosure has simple and quick operation process, mild digestion process, simple required instruments and equipment, and low cost; the obtained nucleus pulposus cells have a large amount of cells, a high survival rate, and are easier to use. It adheres to the wall in the culture flask, and has a good proliferation rate and cell morphology, and the cell activity is strong.

Description

technical field [0001] The present disclosure relates to a preparation method of primary nucleus pulposus cells. Background technique [0002] The disclosure of information in this Background section is only for enhancement of understanding of the general background of the disclosure and should not necessarily be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person of ordinary skill in the art. [0003] Low back pain is the most common disease in orthopaedics. It not only causes mental pain to patients, but also causes huge economic burden. However, there is still no effective treatment in clinical practice. The intervertebral disc is the largest avascular organ in the human body. Studies have reported that about 40% of patients' low back pain is caused by intervertebral disc degeneration, and the most obvious changes are found in the nucleus pulposus. The current clinical treatment includes non-surgical t...

AI Technical Summary

Problems solved by technology

Although patients with lumbar disc herniation are relatively common, they are older, the nucleus pulposus tissue in the intervertebral disc is aging and the number of nucleus pulposus cells is small, and it is difficult to culture in vitro

However, the traditional primary culture of nucleus pulposus cells is cumbersome, has a high contamination rate, and is prone to failure.

Moreover, the primary nucleus pulposus cells are generally cultured in vitro after 3-4 passages, and the cell morphology gradually changes, which is not conducive to the needs of scientific research. In order to ensure the reliability of the experimental results, cells that have been passed 1-2 times are usually used. nucleus pulposus

[0005] In the prior art, the method for isolating primary cells of the nucleus pulposus is commonly used to digest with trypsin and / or collagenase. Since the cell membrane itself contains protein, the longer the digestion, the enzyme digestion solution can hydrolyze this type of protein. After the protein is hydrolyzed, the cell membrane of the primary nucleus pulposus cells ruptures, which seriously damages the integrity of the primary nucleus pulposus cells, thereby affecting the viability of the isolated primary nucleus pulposus cells and subsequent proliferation and culture of the primary nucleus pulposus cells; and , as the digestion time prolongs, it will also damage the respiratory enzymes of the cells, thereby affecting the metabolism of the cells

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