Application of bifidobacterium longum BL21 and microbial inoculum containing bifidobacterium longum BL21 in preparation of products for preventing, relieving or treating colorectal cancer
A technology of Bifidobacterium longum and colorectal cancer, which is applied in the field of microbiology to achieve the effect of improving the micro-ecological environment, good acid and bile salt resistance, and ensuring balance
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Embodiment 1
[0081] Tolerance analysis of Bifidobacterium longum BL21 to simulated gastrointestinal fluid
[0082] (1) Tolerance of Bifidobacterium longum BL21 to simulated gastric juice
[0083] Bifidobacterium longum BL21 was inoculated into MRS medium at 1%, cultured at 37°C for 16 hours, and centrifuged at 8000 rpm for 5 minutes to collect the bacteria. The bacteria were resuspended in 9 mL of artificial simulated gastric juice with pH 2.0, 2.5 and 3.0 (MRS medium containing 1% pepsin, pH 2.0, 2.5 and 3.0) and left anaerobically at 37°C. Culture for 3h, take samples after 0h and 3h of treatment respectively, measure the number of viable bacteria by pouring culture method and calculate the survival rate, the formula is as follows:
[0084] Survival rate (%)=number of viable bacteria after treatment for 3h / initial number of viable bacteria×100%.
[0085] The test results are shown in Table 1.
[0086] Table 1 Tolerance of Bifidobacterium longum BL21 to simulated gastric juice
[0087...
Embodiment 2
[0097] Adhesion of Bifidobacterium longum BL21 to HT-29 cells in vitro
[0098] 1. HT-29 cell culture passage
[0099] Take out the cryovial containing HT-29 cells from the ultra-low temperature freezer, immediately put it into a constant temperature water bath at 37°C, shake it slightly to make it thaw quickly within 1-1.5min, take out the cryovial, and rinse it with 75% alcohol. After disinfection, put it in the ultra-clean bench. Aspirate the cell suspension into a centrifuge tube with a pipette, add 2mL RPMI 1640 medium (containing 10% double antibody, 1% fetal bovine serum), gently pipette and mix, centrifuge at 1080r / min for 5min, carefully aspirate and discard the cell suspension. Then add 3mL RPMI 1640 medium (containing 10% double antibody, 1% fetal bovine serum), resuspend the cells by gently pipetting, suck the cell suspension into a 25mL culture flask with a pipette, and then add RPMI1640 Culture medium to a total volume of 5 mL, then place at 37 °C, 5% CO 2 Aft...
Embodiment 3
[0108] Inhibitory ability of Bifidobacterium longum BL21 on HT-29 cells
[0109] After the HT-29 cells were cultured and passaged according to the method of Example 2, the cells in log phase were collected, and after digestion with trypsin-EDTA, 4 mL of culture medium was added to terminate the digestion, the cells were resuspended by pipetting, and the cells were counted to estimate the number of cells. , and then adjust the cell concentration to 1 × 10 with the medium 5 / mL, take 100 μL of cell suspension and inoculate it in the middle well of a 96-well plate, and place it at 37°C, 5% CO. 2 The cells were cultured in a cell incubator. After the cells had completely adhered to the wall, the old culture medium was discarded, and 100 μL of RPMI 1640 medium (antibody-free) was added for grouping experiments. A total of 6 groups were set up in the experiment, including Bifidobacterium longum BL21 group, Lactobacillus rhamnosus LRa05 group, BL21+LRa05 combined group, Bifidobacter...
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