【摘要】：目的探索采用Cell Counting Kit-8(CCK-8)試劑檢測新鮮臍帶不同大小與數(shù)量組織中細(xì)胞活性,選用合適的組織塊大小與數(shù)量誘導(dǎo)培養(yǎng)間充質(zhì)干細(xì)胞,以獲得最佳的細(xì)胞數(shù)量與細(xì)胞活性。方法臨床獲取新鮮臍帶加工處理剪切成1/8組織塊大小,重量為(0.045±0.001)g,采用CCK-8檢測試劑盒檢測新鮮臍帶組織塊活性,對CCK-8試劑的劑量、加入試劑后孵育時間、臍帶組織塊剪切大小與數(shù)量等做了比較與分析,并在6孔板中加入3塊不同臍帶組織塊大小(1/2組、1/4組、1/8組)誘導(dǎo)培養(yǎng)間充質(zhì)干細(xì)胞,對3組生成的細(xì)胞數(shù)量和細(xì)胞活性做了實(shí)驗(yàn)研究。結(jié)果新鮮臍帶組織塊剪切成厚度為0.5 cm左右環(huán)形組織塊的1/8組織塊大小,重量為(0.045±0.001)g,24孔板內(nèi)臍帶組織放置3塊,CCK-8試劑濃度為15%,于37℃下孵育7-8 h為檢測新鮮臍帶組織塊活性的最佳檢測條件;6孔板中加入3塊不同臍帶組織塊大小(1/2組、1/4組、1/8組)誘導(dǎo)培養(yǎng)間充質(zhì)干細(xì)胞,3組中培養(yǎng)生成的細(xì)胞活性無明顯差異(P0.05)。但3組中培養(yǎng)生成的細(xì)胞數(shù)量,其中1/8組與1/2組或1/4組相比,差異有統(tǒng)計學(xué)意義(P0.01),且每克組織塊生成的細(xì)胞數(shù)量最多。結(jié)論 CCK-8試劑盒可用于準(zhǔn)確地評估新鮮臍帶組織中細(xì)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 鹵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 鹵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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