纳米细菌培养及其对人脐静脉内皮细胞生物学形状的影响和机制探讨

发布时间：2018-08-19 19:47

【摘要】：第一部分 纳米细菌的培养、收获与鉴定 目的：探讨纳米细菌的培养和收获方法，检测培养过程中物质消耗情况和生长速度变化，鉴定培养后的纳米细菌。 方法：在纳米细菌的培养过程中，寻找培养的适宜条件，防污染及收获方法。用全自动生化仪动态检测培养上清液中电解质和葡萄糖的变化情况。使用酶标仪动态检测纳米细菌的吸光度值，间接了解其生长速度。通过使用纳米细菌单克隆抗体进行免疫荧光试验鉴定培养的纳米细菌和利用电镜进一步观察其形状。 结果：纳米细菌适宜的培养条件是：使用细胞培养方法，含10％FBS的完全培养基，加入纳米细菌后，在培养箱中培养，培养箱设置为37℃，5％CO2和95％空气培养。培养过程中采用一系列措施可有效地避免纳米细菌污染。培养的纳米细菌用细胞刮子刮离瓶底，吸入离心管中，4℃离心45分钟，将细菌粉末-20℃保存，可以较好地收获纳米细菌。培养过程中，上清液中的K、Na、Cl、Mg及葡萄糖的浓度无明显变化(P＞0.05)，而Ca、P进行性减少，每周检测结果比较差异有显著性(P＜0.05)。纳米细菌吸光度动态检测结果显示，其吸光度值逐渐增加，每周检测结果比较差异有显著性(P＜0.05)。使用免疫荧光的方法检测时，，纳米细菌可发出绿色荧光，透视电镜下测量纳米细菌大小为80—300nm。
[Abstract]:The first part of the culture, harvest and identification of nanobacteria purpose: to explore the cultivation and harvesting methods of nanobacteria, The changes of substance consumption and growth rate during culture were detected to identify the cultured nanobacteria. Methods: in the process of nanobacteria culture, the suitable conditions, pollution prevention and harvest methods were found. The changes of electrolyte and glucose in culture supernatant were detected by automatic biochemical instrument. The absorbance value of nanobacteria was dynamically detected by enzyme-labeled instrument, and the growth rate of nanobacteria was indirectly understood. The cultured nanobacteria were identified by immunofluorescence assay using monoclonal antibodies against nanobacteria and their shapes were observed by electron microscope. Results: the suitable culture conditions for nanobacteria were as follows: using cell culture method, complete culture medium containing 10%FBS, adding nanobacteria, cultured in incubator, the incubator was set at 37 鈩

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