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Non-cell-derived polypeptide-sensitized DC-CIK cells, and construction method and application thereof
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A technology of DC-CIK and construction method, which is applied in the direction of animal cells, vertebrate cells, cell culture active agents, etc., and can solve the problems of unclear specific antigen and limited anti-tumor effect
Active Publication Date: 2018-07-06
SHANGHAI UNIV +1
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However, due to the lack of specific antigens in many tumors or the lack of specific antigens, the in vivo anti-tumor effect of DC-CIK cells sensitized by tumor antigens is limited.
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Embodiment 1
[0061]Embodiment 1: the culture of A1-DC-CIK cell (can refer to figure 1 shown)
[0064] 2) After centrifugation, carefully draw the upper layer liquid with a pipette gun, stop when it is about 0.5 cm away from the buffy coat layer containing mononuclear cells, and discard the upper layer liquid.
[0066] 4) Discard the supernatant, resuspend the cells with 5-10ml PBS, blow and wash gently with a pipette gun, and centrifuge at 200-300g for 5-10min.
[0067] 5) Repeat 4), discard the supernatant, resuspend the cells with RPMI-1640 complete medium contai...
Embodiment 2
[0092] Embodiment 2: the culture of HCBP1-DC-CIK cell (see figure 1 shown)
[0095] 2) After centrifugation, carefully draw the upper layer liquid with a pipette gun, stop when it is about 0.5 cm away from the buffy coat layer containing mononuclear cells, and discard the upper layer liquid.
[0096] 3) Carefully transfer the buffy coat to a new centrifuge tube with a pipette gun, add an appropriate amount of PBS to the centrifuge tube, and add more liquid than the buffy coat. Centrifuge at 200-300g for 5-10 minutes.
[0097] 4) Discard the supernatant, resuspend the cells with 5-10ml PBS, blow and wash gently with a pipette gun, and centrifuge at 200-300g for 5-10min.
[0098] 5) Repeat 4), discard the supernatant, resuspend the cells with RPMI-1640 complete medium containing ...
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Abstract
The invention discloses non-cell-derived polypeptide-sensitized DC-CIK cells, and a construction method and application thereof. In a plurality of typical embodiments, the construction method comprises the following steps: acquiring peripheral blood, and carrying out isolating to obtain mononuclear cells; isolating mononuclear cells to obtain monocytes and lymphocytes; inducing the differentiationof the monocytes into mature polypeptide-sensitized DC; inducing the differentiation of the lymphocytes into CIK cells; and subjecting the DC and CIK cells to co-culture so as to obtain the polypeptide-sensitized DC-CIK cells. The DC-CIK cells prepared in the invention have high proliferation activity, can specifically kill tumors and are high in tumoricidal activity; and the construction methodhas low requirements on equipment, and is simple in process, easily controllable in conditions and suitable for large-scale implementation.
Description
technical field [0001] The invention particularly relates to a DC-CIK cell sensitized by a non-cell derived polypeptide, its construction method and application, and belongs to the field of biotechnology. Background technique [0002] Cytokine-induced killer cells (CIK) originally refer to a type of CD3 that exists in the peripheral blood of normal people + CD56 + The normal content of T lymphocytes is about 1%-5%. It is also distributed in the liver, and the distribution is the most, followed by peripheral blood, but the existence of this cell has not been found in human bone marrow cells so far. In 1991, Schmidt-wolf et al reported for the first time that adding a series of cytokines to human peripheral blood lymphocytes in vitro could induce a large number of immune-killing cells, namely CIK cells. CIK cells express both CD3 and CD56 double-positive molecules on the surface, so they have both the strong anti-tumor activity of T lymphocytes and the non-major histocompat...
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