CCK-8检测新鲜脐带组织中细胞活性的初步研究

发布时间：2018-12-17 03:07

【摘要】：目的探索采用Cell Counting Kit-8(CCK-8)试剂检测新鲜脐带不同大小与数量组织中细胞活性,选用合适的组织块大小与数量诱导培养间充质干细胞,以获得最佳的细胞数量与细胞活性。方法临床获取新鲜脐带加工处理剪切成1/8组织块大小,重量为(0.045±0.001)g,采用CCK-8检测试剂盒检测新鲜脐带组织块活性,对CCK-8试剂的剂量、加入试剂后孵育时间、脐带组织块剪切大小与数量等做了比较与分析,并在6孔板中加入3块不同脐带组织块大小(1/2组、1/4组、1/8组)诱导培养间充质干细胞,对3组生成的细胞数量和细胞活性做了实验研究。结果新鲜脐带组织块剪切成厚度为0.5 cm左右环形组织块的1/8组织块大小,重量为(0.045±0.001)g,24孔板内脐带组织放置3块,CCK-8试剂浓度为15%,于37℃下孵育7-8 h为检测新鲜脐带组织块活性的最佳检测条件;6孔板中加入3块不同脐带组织块大小(1/2组、1/4组、1/8组)诱导培养间充质干细胞,3组中培养生成的细胞活性无明显差异(P0.05)。但3组中培养生成的细胞数量,其中1/8组与1/2组或1/4组相比,差异有统计学意义(P0.01),且每克组织块生成的细胞数量最多。结论 CCK-8试剂盒可用于准确地评估新鲜脐带组织中细胞活性。
[Abstract]:Objective to explore the use of Cell Counting Kit-8 (CCK-8) reagent to detect the cell activity in different size and quantity of fresh umbilical cord tissues, and to select the appropriate tissue block size and number to induce the culture of mesenchymal stem cells. To obtain the best cell number and cell activity. Methods the fresh umbilical cord was cut into 1 / 8 tissue mass and the weight was (0.045 卤0.001) g. The CCK-8 assay kit was used to detect the activity of the fresh cord tissue. The dose of CCK-8 reagent was measured. The incubation time, shear size and quantity of umbilical cord tissue mass were compared and analyzed after incubating with reagents, and three different umbilical cord tissue blocks were added to the 6-well plate (1 / 2 group, 1 / 4 group, 1 / 8 group) to induce the culture of mesenchymal stem cells. The number and activity of three groups of cells were studied. Results the fresh umbilical cord tissue mass was cut into 1 / 8 of the circular tissue mass about 0.5 cm in thickness, and the weight was (0.045 卤0.001) g ~ (24) hole in the umbilical cord tissue. The concentration of CCK-8 reagent was 15%. Incubation at 37 鈩，

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