Application of bacteroides fragilis or Akkermansia muciniphila to preparation of drug for preventing or treating tumors
A technology of Bacteroides fragilis and myxobacteria, applied in the field of biomedicine
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Embodiment 1
[0042] Embodiment 1 Bacteroides fragilis culture
[0043] training method
[0044] Step 1: Take a freeze-dried and preserved strain of Bacteroides fragilis (purchased from ATCC official website), add 200 μL TSB medium, reconstitute, draw 20 μL, streak the blood plate, and pump air from the gas control system of the anaerobic tank Then in a biochemical incubator at 37°C, anaerobic culture for 48h;
[0045] Step 2: pick a single clone colony into 10mL TSB medium, 37°C, anaerobic culture for 12h;
[0046] Step 3: Take a bottle of 500mL TSB medium, insert 1% (v / v) strains, and culture anaerobically at 37°C for 48 hours;
[0047] Step 4: Take the bacterial solution and centrifuge at 6000rpm for 10min. Wash twice with normal saline, and finally reconstitute the sludge with normal saline for use and count the viable bacteria.
Embodiment 2
[0048] Embodiment 2 Ackerman's myxobacteria culture
[0049] training method
[0050] Step 1: Take a freeze-dried strain of Akkermansia muciniphila (purchased from ATCC official website), add 200 μL TSB medium, reconstitute, draw 20 μL, streak the blood plate, anaerobic tank gas control system After pumping, incubate in a biochemical incubator at 37°C for 48 hours under anaerobic conditions;
[0051] Step 2: pick a single clone colony into 10mL TSB medium, 37°C, anaerobic culture for 12h;
[0052] Step 3: Take a bottle of 500mL TSB medium, insert 1% (v / v) strains, and culture anaerobically at 37°C for 48 hours;
[0053] Step 4: Take the bacterial solution and centrifuge at 6000rpm for 10min. Wash twice with normal saline, and finally reconstitute the sludge with normal saline for use and count the viable bacteria.
Embodiment 3
[0054] Example 3 Bacteroides fragilis promotes the infiltration and / or accumulation of CD8+ T cells in the tumor microenvironment and the effect of treating tumors
[0055] figure 1 Schematic diagram of the experimental flow for detecting the accumulation of CD8+ T cells in the tumor microenvironment and the therapeutic effect of B. fragilis and inactivated B. fragilis.
[0056] 1. Training method
[0057] The culture method of Bacteroides fragilis is the same as in Example 1.
[0058] 2. Sample preparation
[0059] 1) Preparation of live Bacteroides fragilis ZY-312 cells
[0060] Step 1: Take a freeze-dried strain (purchased through commercial channels), add 200ul freeze-dried culture medium, redissolve, absorb 20ul, draw a line on the blood plate, and perform biochemical culture after the gas control system of the anaerobic tank is pumped In the box at 37°C, anaerobic culture for 48h;
[0061] Step 2: Pick a monoclonal colony and insert it into 10mL TSB medium, and cult...
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