Effect of optimized collagenase digestion on isolated and cultured nucleus pulposus cells in degenerated intervertebral discs.
Effect of optimized collagenase digestion on isolated and cultured nucleus pulposus cells in degenerated intervertebral discs.
复制标题
优化的胶原酶消化对椎间盘退化和培养的核细胞细胞的影响。
DOI：
10.1097/md.0000000000012977
复制
发表时间：
2018-11
期刊：
影响因子：
1.6
通讯作者：
Xu DF
中科院分区：
文献类型：
--
作者：
Feng X;Liu L;Yu BQ;Huang JM;Gu LD;Xu DF
本研究旨在通过观察不同浓度的I型和II型胶原酶对退变椎间盘髓核细胞的消化作用，探索胶原酶消化髓核细胞的最佳方法。从18例椎间盘突出患者的椎间盘标本中提取髓核蛋白，用Ⅰ型和Ⅱ型胶原酶消化，并以不同浓度进行分离或组合。在消化后4、8、16和24小时，使用倒置显微镜计数NP细胞，并通过台盼蓝染色测定活性。同时观察NP细胞的生长情况。与胶原酶组合的NP细胞的量大于以相同浓度分离的NP细胞的量。在4小时和8小时的组合胶原酶中，浓度越高，NP细胞的量变得越大。极低浓度的胶原酶作用16和24 h后细胞数量增加，其活性保持在较高水平。采用极低浓度的I型和II型胶原酶联合优化消化，既节省酶，又对NP细胞损伤小，更适合分离培养的NP细胞。
This study aims to explore the optimized digestive method of collagenase to nucleus pulposus (NP) cells by observing the digestive effects of type I and II collagenase in different concentrations to NP in degenerated intervetebral discs. NP were collected from 18 human herniated intervertebral disc samples, and digested by type I and II collagenase, which were separated or combined in different concentrations. NP cells were counted using an inverted microscope, and the activities were determined by trypan blue staining at 4, 8, 16, and 24 hours after digestion. The growth of NP cells was also observed. The amount of NP cells with combined collagenases was greater than that separated in an identical concentration. With the combined collagenases at 4 and 8 hours, the higher concentration, the greater the amount of NP cells became. The amount of cells in extremely low concentrations of collagenase increased after 16 and 24 hours, and its activities remained at a higher level. The optimized digestion of extremely low concentrations of type I and II collagenase combined could save enzymes, was less harmful to NP cells, and was more adapted to separated and cultured NP cells.