【摘要】：生命科學迎來了繁榮發(fā)展的時代。從生物石油到藥品、醫(yī)療,無一不和生命科學息息相關。細胞株作為生命科學研究的重要材料顯得尤為重要。無污染的細胞株是得到可靠實驗數(shù)據(jù)的前提。作為國家級細胞株保藏中心,監(jiān)測細胞株的質(zhì)量一直是本細胞庫重點工作之一。細胞株的微生物污染源一般有細菌、真菌、支原體。細菌和真菌污染細胞培養(yǎng)物時導致培養(yǎng)基變色,鏡下可以看到菌體。但細胞株感染支原后培養(yǎng)基通常不會有顏色變化,更不能在普通顯微鏡下觀察到。支原體會影響培養(yǎng)細胞的各項參數(shù),甚至導致細胞株死亡,因此建立細胞庫支原體檢測體系以及支原體去除方法非常必要。本研究結(jié)合本庫自身情況選用最易操作的三種方法:DNA染色、直接培養(yǎng)法、PCR,對本細胞庫保藏的常用的100種細胞株進行支原體檢測。根據(jù)各不同方法的優(yōu)缺點建立適用于本細胞庫的支原體檢測流程體系:入庫前PCR試劑盒預檢、入庫后三項全檢、日常兩項監(jiān)控、供應中自建PCR檢測。論文在有支原體污染的細胞株中選取珍貴、稀有、且國內(nèi)研究者急需求的細胞株為研究對象,同時使用BM-Cyclin和MC-210兩種藥劑去除支原體。在支原體去除后的一周、一個月、一年的時間段里,重復檢測支原體污染,用以評價兩種藥劑效果和優(yōu)缺點。通過對比,最終確定BM-Cyclin為細胞庫去除支原體污染的首選藥劑。
[Abstract]:Life science ushered in an era of prosperity and development. Everything from biooil to medicine is closely related to life sciences. As an important material in life science research, cell line is very important. Non-polluting cell lines are a prerequisite for reliable experimental data. As a national cell line preservation center, monitoring the quality of cell line has been one of the key tasks of the cell bank. Bacteria, fungi, and mycoplasma are the main sources of microbial pollution in cell lines. Bacteria and fungi contaminate the cell culture resulting in discoloration of the medium, which can be seen under the microscope. However, the culture medium did not change in color after infection, and could not be observed under ordinary microscope. Mycoplasma can affect the parameters of cultured cells and even lead to cell line death. Therefore, it is necessary to establish the detection system of mycoplasma and the removal method of mycoplasma. In this study, we selected the three most convenient methods: DNA staining, direct culture, and PCR, to detect mycoplasma of 100 common cell lines preserved in our library. According to the merits and demerits of different methods, the flow system of mycoplasma detection suitable for this cell library was established: pre-examination of PCR kit before storage, three full tests after entering the library, two daily monitoring items, and self-built PCR detection in supply. In this paper, the rare and precious cell lines contaminated by mycoplasma were selected as the research objects. BM-Cyclin and MC-210 were used to remove mycoplasma at the same time. Mycoplasma contamination was repeatedly detected one week, one month and one year after mycoplasma was removed to evaluate the efficacy, advantages and disadvantages of the two agents. By comparison, BM-Cyclin was determined as the first choice for removing mycoplasma contamination from cell banks.
【學位授予單位】：上海交通大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R446;R375

【參考文獻】

相關期刊論文 前4條

1 吳清壇;蒲榮;;固體與液體培養(yǎng)法在鑒別支原體感染的差異性研究[J];中國醫(yī)藥導報;2011年06期

2 曹忠玲;;大規(guī)模細胞培養(yǎng)中控制污染的舉措[J];中國獸藥雜志;2010年02期

3 杜金玲;王丹娜;石磊;呂超超;王吉瑋;孟姍姍;劉長輝;盧會英;趙亞榮;;生物制品中支原體污染及清除方法的研究進展[J];中國獸藥雜志;2012年02期

4 甘一迪;王銀銀;任芳麗;張旭;田碩;張誠;;培養(yǎng)細胞污染支原體的PCR檢測方法的建立及應用[J];中國醫(yī)藥生物技術;2011年04期

，

本文編號：2274333