Method for in vitro culture of high-proliferation and high-mortality NK cells
A technology of NK cells and cell culture, applied in the field of cellular immunity, to achieve the effect of high proliferation and high lethality
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[0022] Example 1: The method for expanding NK cells in vitro provided by the present invention
[0023] A coating solution containing anti-human CD3 and anti-human CD16 monoclonal antibodies was added to the cell culture flask in advance, and incubated for 1 hour at room temperature. The final concentration of the anti-human CD3 and anti-human CD16 monoclonal antibodies were both 0.15 mg / ml.
[0024] Collect the patient’s peripheral blood mononuclear cells by a hematology separator, transfer the collected blood sample to a centrifuge tube; centrifuge at 700g for 10 minutes, absorb the upper layer of plasma for use in culture; use 0.9% saline blood sample to restore the sample to its original volume and mix; The diluted blood was slowly added to Ficoll, and centrifuged at 900g for 20 minutes; the milky white mononuclear cell layer at the interface of the separated liquid was drawn; washed twice by centrifugation and counted; resuspended the PBMC with serum-free medium and adjusted th...
Example Embodiment
[0027] Example 2: The method for expanding NK cells in vitro provided by the present invention
[0028] Preliminarily add a coating solution containing anti-human CD3 and anti-human CD16 monoclonal antibodies to the cell culture flask, and incubate at room temperature for 6 hours. The final concentration of the anti-human CD3 and anti-human CD16 monoclonal antibodies is 0.01 mg / ml.
[0029] Collect the patient’s peripheral blood mononuclear cells by a hematology separator, transfer the collected blood sample to a centrifuge tube; centrifuge at 700g for 10 minutes, absorb the upper layer of plasma for use in culture; use 0.9% saline blood sample to restore the sample to its original volume and mix; The diluted blood was slowly added to Ficoll, and centrifuged at 900g for 20 minutes; the milky white mononuclear cell layer at the interface of the separated liquid was drawn; washed twice by centrifugation and counted; resuspended the PBMC with serum-free medium and adjusted the cell con...
Example Embodiment
[0032] Example 3: The method for expanding NK cells in vitro provided by the present invention
[0033] Preliminarily add a coating solution containing anti-human CD3 and anti-human CD16 monoclonal antibodies to the cell culture flask, and incubate at room temperature for 0.5 h. The final concentration of the anti-human CD3 and anti-human CD16 monoclonal antibodies are both 3.0 mg / ml.
[0034] Collect the patient’s peripheral blood mononuclear cells by a hematology separator, transfer the collected blood sample to a centrifuge tube; centrifuge at 700g for 10 minutes, absorb the upper layer of plasma for use in culture; use 0.9% saline blood sample to restore the sample to its original volume and mix; The diluted blood was slowly added to Ficoll, and centrifuged at 900g for 20 minutes; the milky white mononuclear cell layer at the interface of the separated liquid was drawn; washed twice by centrifugation and counted; resuspended the PBMC with serum-free medium and adjusted the cell ...
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