CAJ | 학술논문

目的链亲和素标记的细胞因子锚定MB49细胞和成瘤小鼠膀胱,探讨锚定率、锚定量与时间的关系.方法 MB49细胞与链亲和素标记的绿色荧光蛋白(SA-GFP)体外锚定后,在荧光显微镜下观察;建立小鼠原位表浅膀胱癌模型后,实验组膀胱灌注锚定链亲和素标记的粒细胞-巨噬细胞集落刺激因子(SA-GM-CSF);对照组小鼠不锚定直接膀胱灌注SA-GM-CSF.取膀胱作冰冻切片免疫组织化学分析.结果 MB49细胞基本被SA-GFP锚定,锚定率为98.78%;冰冻切片免疫组织化学显示实验组膀胱黏膜有阳性显色,以第1天最浓,以后逐日变淡,至第7天仍可见较明显染色;对照组膀胱黏膜无阳性显色.结论链亲和素标记的细胞因子可以锚定在细胞表面,且锚定效率高、保留时间长.
목적 련친화소표기적세포인자묘정MB49세포화성류소서방광,탐토묘정솔、묘정량여시간적관계.방법 MB49세포여련친화소표기적록색형광단백(SA-GFP)체외묘정후,재형광현미경하관찰;건립소서원위표천방광암모형후,실험조방광관주묘정련친화소표기적립세포-거서세포집락자격인자(SA-GM-CSF);대조조소서불묘정직접방광관주SA-GM-CSF.취방광작빙동절편면역조직화학분석.결과 MB49세포기본피SA-GFP묘정,묘정솔위98.78%;빙동절편면역조직화학현시실험조방광점막유양성현색,이제1천최농,이후축일변담,지제7천잉가견교명현염색;대조조방광점막무양성현색.결론 련친화소표기적세포인자가이묘정재세포표면,차묘정효솔고、보류시간장.
Objective Streptavidin (SA)-tagged cytokine was anchored to MB49 cells in vitro and biotinylated mucosal surface of bladder wall in vivo. The anchoring rate was calculated and the effects of anchoring and the relation between anchoring effect and time were observed. Methods Streptavidin ( SA) tagged green fluorescent protein (SA-GFP) was anchored to MB49 cells in vitro and observed under an fluorescence microscope. Female C57BL/6 mice with superficial bladder cancer were intravesically instilled with NHS-PE04-Biotin, PBS, SA-GM-CSF as SA-GM-CSF group, and PBS, SA-GM-CSF as PBS group.All the mice were killed to obtain their snap-frozen bladders for cryosectioning, and then immunohistochemistry analysis with biotin-conjugated HRP and DAB display liquid kit. Results SA-GFP was anchored to the surface of MB49 cells under the fluorescence microscopy, and the anchoring rate was 98.78%.SA-GM-CSF was detected on the biotinylated mucosal surface of bladder wall up to 7 days after its intravesical immobilization in SA-GM-CSF group, but no SA-GM-CSF was found in PBS group. Conclusion SA-GFP can be anchored to the surface of MB49 cells, and the anchoring rate was 98. 78%.