Human umbilical cord blood immune cell bank establishing method

Abstract

A human umbilical cord blood immune cell bank establishing method comprises the steps that umbilical cord blood collection and transportation are performed; a mononuclear cell is separated from umbilical cord blood; a CD34+ cell in the mononuclear cell is induced into a DC cell; suspending lymphocyte and the wall-attaching DC cell are separated; a tumor antigen peptide is selected to conduct antigen supporting on the DC cell so as to obtain a mature DC cell; the suspending lymphocyte is cultured to obtain a CIK cell; the mature DC cell is added to the CIK cell to perform large-scale co-culture, then a DC-CIK cell is collected, detection is performed, the cell is put in an immune cell bank for cryopreservation after being qualified, and a cell file is established; a corresponding effect cell type is selected from the immune cell bank according to clinical diagnosis, and retransformation is performed after resuscitaion culture for 24 hours. The immune cell bank established by adopting the method is a set of various tumor antigen peptide effect cells, patients can select appropriate effect cells for resuscitaion and retransformation when needing cell treatment, the time waiting for cell culture is shortened, the best treatment period is mastered, and a treatment effect is improved.

Description

technical field [0001] The invention relates to a method for constructing a cell bank, in particular to a method for constructing an immune cell bank from human umbilical cord blood. Background technique [0002] Immune cells generally refer to all cells participating in the immune response, and specifically refer to lymphocytes that can recognize antigens and generate specific immune responses. Immune cell therapy is to collect human immune cells, under the action of various immune active factors, after in vitro culture, a large number of immune effector cells are expanded, and then reinfused into the body to kill pathogens and cancer cells in blood and tissues. At present, the immune cells used for clinical treatment mainly include dendritic cells (DC), cytokine-induced killer cells (CIK), DC-stimulated CIK cells (DC-CIK), natural killer cells (NK), tumor infiltrating Lymphocytes (TIL), etc. [0003] Immune cell therapy has become the fourth tumor treatment technology fo...

AI Technical Summary

Problems solved by technology

However, there are several unavoidable problems in this treatment method: (1) Due to the isolation and induced culture of mononuclear cells after blood collection, if patients choose immune cell therapy, they have to wait at least two weeks, and it is easy to miss the best treatment period, and Extended hospital stay; (2) Cancer patients, especially advanced cancer patients, have weak constitution and low immune function, and generally cannot tolerate repeated blood draws (such as easily induced ischemic anemia, etc.), so the amount of autologous blood obtained is less, and One-time culture can only be expanded by about 70-80 times, which is difficult to meet the number of cells required for immune cell therapy. Clinicians need to balance cell therapy and repeated blood draws; (3) The quality of the patient's own blood is not high, especially the antigen The number of presenting cells (DC cells are the most efficient antigen-presenting cells) is small, while the number of immunosuppressive cells (Treg cells) is large. The number of immunosuppressive cells is also increasing during induction culture, and will continue to a certain extent. Inhibition of effector cells; (4) Clinical immune cell therapy (DC-CIK) mostly uses the full antigen loading mode (such as CN102861107B), the targeting of the treatment is not strong, and there is a risk of graft-versus-host reaction

However, mononuclear cells, especially induced lymphocytes, are fragile. Due to the toxic effect of DMSO and the mechanical operation during cryopreservation, it is easy to cause changes in the configuration of certain proteins on the immune cell membrane, thereby causing changes in cell subtypes. , leading to a decrease in the secretion of perforin; on the other hand, the cells need to pay attention to the removal of DMSO during the resuscitation treatment. It has been reported that the incidence of adverse events in cell therapy is related to the amount of DMSO reinfusion

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