Ovarian Cancer Stem Cell Separation Culture And Antigen Identification

Objective:Using different methods to separate educational ovarian cancer of the organization and the specimen of ovarian cancer stem cell and by different methods for verification and sure of ovarian cancer stem cell and to the surface of the antigens for testing for the next step to the tumor peptied vaccine research laid the foundation.Methods:(1) Ovarian cancer stem cell culture and separation:Conventional digest, breakdown of the tumor tissues arise with the breakdown of erythrocytes beads satisfactions apparatus satisfactions CD133+ovarian cancer stem cells there, then use the training of ovarian cancer stem cell culture medium ovarian cancer stem cell, the cancer stem cells ascites source of the train, all desire arise ascites breakdown of the breakdown of what, then using stem cells cluture medium training of ovarian cancer stem cell, the ovaries the department and the source of A2780ovcar ovarian cancer stem cell, using stem cells pui (2) Identification of ovarian cancer stem cell:Use of the step of ovarian cancer stem cell to cell CD133streaming the fluorescence detection and immune to the dyeing and cloning stem cells clon in the experiment and generation time, the number of ovarian cancer on a differentiated to renew and trap a;body has the ability to a hundred thousand a cell implant and the number3-4weeks nod scid mouse,right under the formation of differences.(3)Identification of ovarian cancer stem cell surface antigens:Use of quantitative methods of ovarian cancer PCR and what we have ovarian cancer stem cell surface of the antigens differences. the gene is pcAb and may be expressed in the ovary cancer stem cell, there are more representative and more meaningful to the original identification and for the next step to the original form of vaccine research designed to lay foundation for the clinical treatment of ovarian cancer provide the terrorists.Result(1) Organizations and A2780supply of ovarian cancer stem cell Culture by train, look at the cell at about3-5days into a ball and a suspension or grow it, child vaccinations and six Six holes board the train, be a50-100other cells, observed A ball and Clon cell form’s characteristic of stem cells;(2) By MACA satisfactions of cell groups CD133express rate reached82.54%, using stem cells A2780an induction training into a ball, after a streaming cells were expressed by CD133rate is approximately92.69%; by immunization fluorescent and PCR, and differentiation experiment, lymphoma experiments proved that we wait for further the cells of ovarian cancer;(3) Analysis ovarian cancer and ovarian cancer stem cell surface antigen expression Sp17、OY-TES-1、 LAGE、AKAP3、DPPA2etal.The result is the ovarian stem cells of the antigens expression more than ovarian cancer cells.Conclusion:Successful separate of ovarian cancer infection of ovarian cancer stem cell, adopted to stem cells an ovarian cancer stem cell, and adopt a different way from different aspects of the identification of ovarian cancer stem cell of ovarian cancer. Further with ovarian cancer stem cell of the surface of the antigens expressing the comparative analysis, for the next step vaccine research lay a foundation.