Immune cell and preparation and application thereof
A technology of immune cells and preparations, applied in the field of preparations and applications of immune cells, which can solve the problems of short survival time, low lethality, and less effective effect than hematological tumors, and achieve the effect of lasting time and reducing exhaustion
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Embodiment 1
[0093] The preparation method and functional verification of immune cells in this embodiment include the following steps:
[0094] Step 1: Isolation of peripheral blood PBMCs and expansion of CAR-T cells
[0095] Mononuclear cells were isolated from donor peripheral blood, density gradient centrifugation was performed using ficol, and T cells were enriched with a T cell sorting kit (CD3 MicroBeads, human-lyophilized, 130-097-043), using a conjugated anti-CD3 / anti-CD28 magnetic beads activated culture and expansion of T cells; the medium used TexMACS GMP Medium (MiltenyiBiotec, 170-076-309), containing 10% FBS, 2mM L-glutamine, 100IU / ml rhIL2, all cells were placed Cultured in a constant temperature incubator at 37°C with 5% CO2.
[0096] Step 2: Cell Line Culture
[0097] Cell line expressing GPC3: Huh-7 (human liver cancer cell), purchased from ATCC.
[0098] Cell line not expressing GPC3: A549 (human non-small cell lung cancer cells), purchased from ATCC.
[0099] Cells...
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