摘要
目的:用光交联剂SANPAH将不同浓度鼠抗人CD34抗体接枝于经过去细胞和光氧化后的牛颈静脉表面,明确接枝鼠抗人CD34抗体的最佳反应摩尔比和最佳浓度。方法:实验于2006-06/08在中南大学湘雅二医院生物材料实验室完成。按4个不同摩尔比(1∶0,1∶10,1∶20,1∶50),SANPAH和4个不同浓度[0.665μmol/L(100mg/L),1.33μmol/L(200mg/L),6.65μmol/L(1g/L),13.3μmol/L(2g/L)]鼠抗人CD34抗体进行反应后,经紫外线照射光化学接枝于经过去细胞和光氧化后的牛颈静脉上,各组进行快速冰冻切片,滴加FITC标记的羊抗鼠IgG,然后在荧光显微镜下观察不同摩尔比和浓度反应结合的荧光效果,从而初步推断出最佳的反应和结合的摩尔比和浓度。结果:随着鼠抗人CD34抗体的浓度的升高,荧光整体上是越来越强,但是当浓度高于6.65μmol/L时,荧光差别不是很明显;当二者的反应摩尔比为1∶20时,荧光最强。结论:最佳的鼠抗人CD34抗体和SANPAH反映接枝的浓度是6.65μmol/L,最佳的摩尔比是1∶20。
AIM： To investigate the suitable reactive concentration and ratio of photocrosslinking mouse anti-human CD34 onto the decellularized and photooxidated bovine jugular vein （BJV） matrix intima. METHODS： The experiment was conducted in the Biomaterial Laboratory, Xiangya Second Hospital, Central South University from June to August 2006. After reacted with different concentration photochemical crosslinker SANPAH （1 ： 0, 1 ： 10, 1 ： 20, 1 ： 50）, mouse anti-human CD34 [0.665 μ mol/L（100 mg/L）, 1.33μmol/L （200 mg/L）, 6.65 μ mol/L （1 g/L）, 13.3 μ mol/L （2 g/L）] were coated onto decellularized and photooxidated BJV using ultraviolet ray, and cut into pieces. The pieces were rinsed in a shaking machine for 6 hours. The fast frozen sections of the specimens were observed under the fluorescent microscope after having been dropping goat anti-mouse IgG FITC-labeled. The grafting effect was viewed by different fluorescent brightness, so the suitable reactive concentration and ratio were deduced. RESULTS： The higher the concentrations of mouse anti-human CD34 and SANPAH, the brighter the fluorescence was. But when the concentration of them was higher than 6.65μ mol/L, the difference of fluorescent brightness was not so clear. In the same concentration, the fluorescence was the brightest if the reactive mole ratio between them was 1 ： 20. CONCLUSION： The optimal concentration and reactive mole ratio between mouse anti-human CD34 and SANPAH were 6.65μ mol/L and 1 ： 20, respectively.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第2期319-322,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
湖南省科技计划重大专项基金资助项目(04SK1005)~~
