Application of akkermansia muciniphila or prevotella copri in preparation of medicine for increasing gamma delta T cell accumulation in tumour microenvironment and improving anti-tumour immunity function
A technology of tumor microenvironment and Prevotella, applied in the field of biomedicine, to achieve the effect of inhibiting growth
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Embodiment 1
[0042] Embodiment 1 Ackerman's myxobacteria culture
[0043] training method
[0044] Step 1: Take a freeze-dried and preserved Akkermansia muciniphila strain (purchased from ATCC), add 200 μL TSB medium to dissolve it, take 200 μl blood plate to streak, and put it in an incubator under anaerobic conditions for 37 Cultivate for 48 hours at ℃;
[0045] Step 2: Pick a monoclonal colony and dissolve it in 10 ml of TSB medium, and culture it under anaerobic conditions at 37°C for 48 hours;
[0046] Step 3: Take 500 milliliters of TSB culture medium, inoculate strains according to 1% (v / v), and cultivate under anaerobic conditions at 37°C for 48 hours;
[0047] Step 4: Collect the bacterial liquid and centrifuge at 6000rpm for 10 minutes. After washing twice with normal saline, the bacteria sludge was reconstituted and counted viable bacteria.
Embodiment 2
[0048] Embodiment 2 Prevotella culture
[0049] training method
[0050] Step 1: Take a freeze-dried Prevotella copri strain (purchased from ATCC), add 200 μl of LPYG medium to dissolve it, take 200 μl of blood plate, and culture it in an incubator at 37°C for 48 hours under anaerobic conditions. Hour;
[0051] Step 2: Pick a monoclonal colony and dissolve it in 10 ml of TSB medium, and culture it under anaerobic conditions at 37°C for 48 hours;
[0052] Step 3: Take 500 milliliters of PYG culture medium, inoculate strains at 1% (v / v), and culture under anaerobic conditions at 37°C for 48 hours;
[0053] Step 4: Collect the bacterial liquid and centrifuge at 6000rpm for 10 minutes. After washing twice with normal saline, the bacteria sludge was reconstituted and counted viable bacteria.
Embodiment 3
[0054] Example 3 Akkermansia enhances the infiltration of γδT cells in the tumor microenvironment and treats tumor (liver cancer) experiment
[0055] figure 1 Schematic diagram of the experimental flow for testing Akkermansia or inactivated Akkermansia to increase γδ T cell infiltration and therapy in the tumor microenvironment.
[0056] 1. Training method
[0057] Akerman's myxobacteria culture method is the same as above-mentioned embodiment 1.
[0058] 2. Sample preparation
[0059] 1) Preparation of Ackermannian myxobacteria live cells
[0060] Step 1: Take a freeze-dried and preserved Akkermansia muciniphila strain (purchased from ATCC), add 200 μL TSB medium to dissolve it, take 200 μl blood plate to streak, and put it in an incubator under anaerobic conditions for 37 Cultivate for 48 hours at ℃;
[0061] Step 2: Pick a monoclonal colony and dissolve it in 10 ml of TSB medium, and culture it under anaerobic conditions at 37°C for 48 hours;
[0062] Step 3: Take 500...
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