摘要
原发性肝癌(简称肝癌)是我国常见、难治性恶性肿瘤之一,占我国恶性肿瘤死亡率的第2位。手术切除仍为肝癌治疗的首选方法,但其疗效仍欠满意,术后复发是影响其疗效的主要因素,研究和寻找抗肝癌术后复发方法是近年来国内外学者共同关注的课题。本研究取人外周血,分离出B淋巴细胞并激活。应用PEG融合技术,制备了人体激活B细胞和肝癌细胞融合细胞,检测到融合率为28.6%,融合细胞的存活率为95.6%。观察了激活B细胞表面CD20分子和MHC-Ⅱ分子的表达;融合细胞不仅表达肿瘤细胞表面分子,同时也高表达MHC-Ⅱ分子。
细胞融合技术是近几十年来迅速发展起来的一项新兴细胞工程技术。此技术可以培养新的物种、品系或制备新的细胞工程产品,如抗肿瘤疫苗、单克隆抗体等等。目前来说,细胞融合已经是一项比较成熟的技术了。而在九十年代,兴起了一项新技术——MEMS技术。本项目紧密跟随细胞电融合领域内的发展方向,提出并设计加工出了微电极阵列的细胞电融合芯片。在本课题中,我们通过实验成功验证了细胞电融合的电介质电泳以及细胞在交流电场的排队现象。在利用细胞电融合芯片进行融合的实验中,发现人肝癌细胞系7721细胞在芯片中仅有个别的细胞能够实现两两细胞接触并黏附在电极上,既而实现融合。但是芯片的重复性不好,在以后的实验中基本上观察不到任何现象。
本论文还利用鸡红细胞进行了PEG法和细胞电融合芯片方法的对照实验。在PEG法中,研究了不同分子量、不同浓度的PEG对细胞融合率的影响,得到了PEG法的最佳融合条件;在细胞电融合芯片方法中,不断变化细胞密度、电场强度和电融合液等条件,希望能为电融合芯片以后实验研究提供融合条件方面的数据参考。
Hepatocellular carcinoma (HCC) is one of the most freguent malignancies in China. Its mortality takes the second place among malignant tumors in China. Although the surgical excision is considered the most effective therapy for early-stage patients, only a small proportion of patients with an operable primary lesion may transiently benefit from surgical treatment because of a high recurrence rate of cancer recurrence rate of cancer after operation. To study and search for an approach of anti- recurrence of HCC after operation is expected.In this study the purified HepG2 cells and activated B cells from peripheral blood. Hybrid cells were generated by fusion of human hepatocellular carcinoma HepG2 with activated B cells from peripheral blood. The fused propotion was 28.6%, the survival rate of hybrid cells was 95.6%. The hybrid cells was able to express MHC-Ⅱfrom B cells,and tumor cell surface antigen from HepG2 cells.
The cell fusion technology is a newly developing technology which has developed dramatically. The hybrid cells can be cultured to be a new species or a new celluar engineering products, such as monoclonal antibody、tumor vaccine. Now the cell fusion technology is a very mature technology. In 90’s, there is a new technology--MENS technology. This topic follows the development of cell electrofusion closely, producing and designing the microelectrode array cell electrofusion chip. We verifed the dielectrophoresis theory and the queuing phenomenon of cells in AC electric field by experiments succeedly. In the experiments of cell electrofusion chip, we found only a few 7721 cells were able to cell-to-cell queue and fuse. It demonstrated the cell fusing rate was worse. In the following experiments, we can not observe the queuing and fusion phenomenon , it demonstrated the cell fusing rate was worse. So the cell electrofusion chip must be improved.
This paper takes the comparison experiments of chicken RBC fusion by PEG method and cell electrofusion chip method. On one hand, the author studied and researched the effect tion of cell fusion rate by different molecular weight and different concentration of PEG/DMSO, and found the best fusion chemical condition of PEG method. On the other hand, the author took some experiments by change the conditions
引文
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