| Objective:Malignant tumor is the phenomenon of gene mutation in normal cells in human body,which leads to malignant proliferation.There is no obvious symptom at the initial stage of the disease,which is difficult to find.When patients have specific symptoms,they have advanced to an advanced stage.Therefore,the patient’s mor rate is extremely high.Recent studies have shown that with our continuous understanding and research on combined immunotherapy,this treatment method can provide new ideas and methods for the treatment of tumor cells.Among them,cell adoptive immunotherapy is one of the effective methods to effectively treat cancer.CIK cells prepared from cord blood are currently widely used in cellular adoptive immunotherapy.But,there is currently a problem of higher Treg cells,if we compared with peripheral blood produced by the human body.Therefore,we write this article for the purpose of optimizing the preparation process of umbilical cord blood immune cells,reducing the number of regulatory T cells,enhancing the activity of prepared cells and optimizing the preparation process of cells.Firstly,the peripheral blood cells of human body were cultured.When we finished the culture cycle,we carried out flow detection on the cells,we take a look at the two data results together,and then see if we can find out where the current preparation process doesn’t work.Then add some new adjusting factors into it,so that we can get what we originally wanted.Method:The first step is to culture lymphocytes extracted from peripheral blood cells produced by human body for two weeks.After culture,several cell values were detected by flow cytometry,namely CD3~+CD56~+cells,CD3~+CD4~+cells,CD3~+CD8~+cells,Treg cells and CD8~+CD28~+cells.We use the same preparation process to prepare and culture the cells for two weeks,and then after the finish the culture,we also test these cell values,so that we can screen these cell values one by one,and then we can find out which part of this preparation process is wrong and problematic.In order to achieve the goal of optimizing the preparation process of cord blood immune cells,we added cytokines that have an effect on the in duction of immune cells:IL-15,IL-12,and IL-21 during the culture process.We use a new preparation method for cell preparation and cell culture.During the culture process,the cells were subjected to flow cytometry on the 4th,7th,10th,and 14th days of culture,and the changes of various cell indexes under the two different processes were compared in the same time.In addition,in order to investigate the immune ability of immune cells,we tested the viability of the immune cells after one cycle of preparation,and carried out cell counting and image acquisition on the self-proliferation of the cells at 0h,24h,and 48h.Result:1.The modified preparation process is compared with the immune cells obtained by the original method,it contains a high proportion of Treg cells.2.The modified preparation process is compared with the immune cells obtained by the original method,it reduces the number of Treg cells.3.The modified preparation process is compared with the immune cells obtained by the original method,it adds the value of CD3~+CD56~+cells,CD3~+CD4~+cells,and CD28~+CD8~+cells.4.The modified preparation process is compared with the immune cells obtained by the original method,it reduces the number of CD3~+CD8~+cells.5.The self-proliferation ability of immune cells prepared by the optimized preparation process becomes stronger.Conclusion:1.The optimized preparation process reduces the positive ratio of Treg cells,which is closer to the human body’s own peripheral blood lymphocytes.2.The cells prepared by changing the preparation method improve the immune tolerance of immune cells.3.The optimized preparation process improves the self-proliferation ability of cells. |
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