The mast cells from enzymatically dispersed rat skin were incubated in DMEM mediumcontaining 10% fetal bovine serum. 1 day after culture, the various concentrations of substance P (SP),SP+Ca2+ or concanavalin A (Con A...The mast cells from enzymatically dispersed rat skin were incubated in DMEM mediumcontaining 10% fetal bovine serum. 1 day after culture, the various concentrations of substance P (SP),SP+Ca2+ or concanavalin A (Con A) were added into the media to stimulate mast cells for different periods of time. The media were rapidly seperated from the cultured tells using the micropore filter. The contents of histamine in the media were determined by spectrofluorimetry and the release rates of histaminewere caculated. The results showed that SP and Con A could stimulate in vitro mast cells tO release histamine in a time--and dose-dependent pattern, and that the effect of SP was significantly influenced by theconcentration of Ca2+.展开更多
目的:探索在人胆囊癌GBC-SD细胞系中是否存在具有干细胞特性的SP细胞(side population cells),以及SP细胞、非SP细胞和GBC-SD细胞系表达常见干细胞标记物ABCG2、Oct-4、CD34的情况。方法:采用荧光激活细胞分选技术(FACS)分选出人胆囊癌S...目的:探索在人胆囊癌GBC-SD细胞系中是否存在具有干细胞特性的SP细胞(side population cells),以及SP细胞、非SP细胞和GBC-SD细胞系表达常见干细胞标记物ABCG2、Oct-4、CD34的情况。方法:采用荧光激活细胞分选技术(FACS)分选出人胆囊癌SP细胞和非SP细胞,通过RT-PCR、Western blot、流式细胞术以及免疫荧光化学技术检测SP细胞、非SP细胞以及GBC-SD细胞系表达ABCG2、Oct-4和CD34的情况。结果:人胆囊癌GBC-SD细胞系中存在着少量的具有干细胞潜能的SP细胞,其所占的比例为(0.64±0.08)%,SP细胞相对于非SP细胞和GBC-SD细胞,具有更强的体外侵袭力(P<0.05);并且ABCG2在胆囊癌SP细胞中呈现出高表达的状态(89.56±3.86)%,在非SP细胞中几乎不表达(1.32±0.49)%,在未分选胆囊癌细胞系中ABCG2呈弱表达(12.37±1.61)%,P=0.001;Oct-4在三种细胞中的表达分别为:(94.87±1.40)%、(88.16±2.34)%、(90.17±1.61)%,P>0.05;CD34在mRNA水平高表达于非SP细胞和GBC-SD细胞,不表达于SP细胞;在蛋白水平几乎均不表达于这三种细胞,其含量依次为:(1.78±0.51)%、(0.63±0.21)%、(0.96±0_38)%,P>0.05。结论:人胆囊癌GBC-SD细胞系中存在具有干细胞特性的SP细胞,并且具有ABCG2^+Oct-4^+CD34^-的表型特征。展开更多
文摘The mast cells from enzymatically dispersed rat skin were incubated in DMEM mediumcontaining 10% fetal bovine serum. 1 day after culture, the various concentrations of substance P (SP),SP+Ca2+ or concanavalin A (Con A) were added into the media to stimulate mast cells for different periods of time. The media were rapidly seperated from the cultured tells using the micropore filter. The contents of histamine in the media were determined by spectrofluorimetry and the release rates of histaminewere caculated. The results showed that SP and Con A could stimulate in vitro mast cells tO release histamine in a time--and dose-dependent pattern, and that the effect of SP was significantly influenced by theconcentration of Ca2+.
文摘目的:探索在人胆囊癌GBC-SD细胞系中是否存在具有干细胞特性的SP细胞(side population cells),以及SP细胞、非SP细胞和GBC-SD细胞系表达常见干细胞标记物ABCG2、Oct-4、CD34的情况。方法:采用荧光激活细胞分选技术(FACS)分选出人胆囊癌SP细胞和非SP细胞,通过RT-PCR、Western blot、流式细胞术以及免疫荧光化学技术检测SP细胞、非SP细胞以及GBC-SD细胞系表达ABCG2、Oct-4和CD34的情况。结果:人胆囊癌GBC-SD细胞系中存在着少量的具有干细胞潜能的SP细胞,其所占的比例为(0.64±0.08)%,SP细胞相对于非SP细胞和GBC-SD细胞,具有更强的体外侵袭力(P<0.05);并且ABCG2在胆囊癌SP细胞中呈现出高表达的状态(89.56±3.86)%,在非SP细胞中几乎不表达(1.32±0.49)%,在未分选胆囊癌细胞系中ABCG2呈弱表达(12.37±1.61)%,P=0.001;Oct-4在三种细胞中的表达分别为:(94.87±1.40)%、(88.16±2.34)%、(90.17±1.61)%,P>0.05;CD34在mRNA水平高表达于非SP细胞和GBC-SD细胞,不表达于SP细胞;在蛋白水平几乎均不表达于这三种细胞,其含量依次为:(1.78±0.51)%、(0.63±0.21)%、(0.96±0_38)%,P>0.05。结论:人胆囊癌GBC-SD细胞系中存在具有干细胞特性的SP细胞,并且具有ABCG2^+Oct-4^+CD34^-的表型特征。
