论文部分内容阅读
目的观察双丹口服液(SOL)对体外分离培养的SD仔鼠心肌干细胞(CSCs)的作用。方法分离培养CSCs,将培养的第2代CSCs分为对照组及不同浓度1×10-1mol/L、1×10-2mol/L、1×10-3mol/L、1×10-4mol/L、1×10-5mol/L和1×10-6mol/L的SOL组。培养24 h、48 h后用MTT比色法检测SOL对CSCs的影响。将10-2mol/L SOL组的细胞培养24 h后,用流式细胞仪测定c-kit+/CD45-细胞的比率。结果消化后的心肌组织3 d后,可见成纤维样细胞从组织块儿周围爬出,1周后可见小、圆、亮的细胞出现在组织块周围爬出的成纤维细胞层上。传代培养后,倒置显微镜下观察细胞形态呈较均一的梭型,可见CSCs形成的“太阳状”集落。MTT法检测表明,与对照组相比,SOL作用后的CSCs生长增殖迅速,当SOL的浓度超过1×10-3mol/L时,CSCs增殖更为显著(P<0.05)。流式细胞术检测显示,1×10-2mol/L SOL的细胞培养24 h后,c-kit+/CD45-细胞的比率达到0.976%,与对照组相比(0.301%),差异具有统计学意义(P<0.01)。结论利用酶消化可以成功从SD仔鼠心脏组织中分离培养得到c-kit+的CSCs,SOL对CSCs的体外生长和增殖有一定的促进作用。
Objective To observe the effects of Shuang Dan oral liquid (SOL) on isolated rat cardiac stem cells (CSCs) cultured in vitro. Methods CSCs were isolated and cultured, and the second passage CSCs cultured in vitro were divided into control group and different concentrations of 1 × 10-1mol / L, 1 × 10-2mol / L, 1 × 10-3mol / L, 1 × 10-4mol / L , 1 × 10 -5 mol / L, and 1 × 10 -6 mol / L SOL group. After cultured for 24 h and 48 h, the effect of SOL on CSCs was detected by MTT assay. The cells in 10-2mol / L SOL group were cultured for 24 h, and the ratio of c-kit + / CD45- cells was determined by flow cytometry. RESULTS: After 3 days of digestion, the fibroblast-like cells could climb out of the tissue block. After 1 week, small, round and bright cells appeared on the fibroblast layer that climbed out of the tissue block. After subculture, the morphology of the cells was observed to be a more uniform spindle shape under an inverted microscope, and the “sun-like” colonies formed by CSCs were observed. The results of MTT assay showed that compared with the control group, the proliferation of CSCs after SOL treatment was rapid. When the concentration of SOL exceeded 1 × 10-3 mol / L, the proliferation of CSCs was more significant (P <0.05). Flow cytometry showed that the ratio of c-kit + / CD45- cells reached 0.976% at 1 × 10-2 mol / L SOL for 24 h, which was significantly lower than that of the control group (0.301%) (P <0.01). Conclusions C-kit + CSCs can be successfully isolated and cultured from cardiac tissue of SD offspring by enzymatic digestion. SOL can promote the growth and proliferation of CSCs in vitro.