Large-scale Production Through Microcarrier Cultivation Followed By Anti-tumor Study Of TALL-104 Cells

Objection : Cancer is always a huge problem for human. Now TALL-104 cell line is one of the new way to cure cancer through cell immunotherapy. This cell line which is a cytotoxic T cell attained from a baby who had Acute lymphocytic leukemia T cells. TALL-104 cell line can kill many tumors(MCF-7, primary malignant brain tumor and so on). There are too many advantages of the cell: 1. No MHC restriction. So it can be used for allogenous treatment and can produce a large number of cells in the factory; 2. It is a cell line, so it can be proliferate without limitation; 3. According to the known clinical trials, in general doses there is almost no allergic reaction; 4. Killing the cancer cells does not need IL-2 (leading to serious side effects). TALL-104 cells is a very cell therapy drug, but now most labs culture the cell in the plate or cell factory. It is very hard to meet the requirements of clinical use. The operation is also difficult and it is hard to control the quality of the production. In the experiment, we use spinner bottle to culture TALL-104 with microcarrier. We also test that the top concentration of the cell with different mediums. The cell line is clinging to the microcarrier, so digestion is a must to obtain the production. Through the experiment, we know the better conditions to get production.Methods: TALL-104 cell was seeded in the plate, in spinner bottle without microcarrier and spinner bottle with microcarrier (Cytodex 3). Concentration of glucose was determined on Nova Bioprofile 100+ and the correspond cells number on the culture process were determined through hemacytometry. We cultured the cell with different medium to compare their effect to Cell growth power curve. Digestion with trypsin is a must to obtain cells after culture, so the best condition for trypsin is very important. We tested different concentration of trypsin and different digestion time to find the best condition to collect cells. MTT was used to compare whether there is different between the TALL-104 cultured with microcarrier and that cultured in the plate.Results: Tall-104 can propagate with 100 IU/mL of IL-2 and in the stirring bottle the cell can grow well on the microcarrier; and the cell cultured in the bioreactor have no different with the cell in the plate to kill the cancer cell; in the factory the best choice is to use the tryspin with EDTA to digest the cell 3 minutes.Conclusion: TALL-104 could use microcarrier to obtain large-scale production in the factory. 1640 medium was better than DMEM to culture TALL-104. In the factory, we recommended 0.25% trypsin with EDTA to digest cell with 3 min.