Method for preparing microbial ecological agent for treating infertility
A micro-ecological preparation and infertility technology, applied in the field of microorganisms, can solve problems such as the influence of sperm cell motility, and achieve the effects of good promotion prospects, low recurrence probability and significant treatment effect.
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Embodiment 1
[0032] A preparation method for probiotics for infertility treatment, comprising the following steps:
[0033] A. Culture and isolation of bacterial flora
[0034] (1) Preparation of medium: LB basal medium, MRS solid medium, BHI solid medium, BHI+5% skim milk medium, BSM solid medium;
[0035] (2) Take out the vaginal secretion sample preserved with glycerin from the -80 degree refrigerator, and after it melts, take an appropriate amount into a sterile 1.5ml EP tube, and use sterile PBS according to 10 1 —10 6 Doubling dilution, MRS, LB, BSM by 10 2 、10 4 Dilute BHI, BHI+5% skim milk by 10 3 、10 5 50 μL of the diluted solution was spread on solid medium, and cultured for 48 hours under anaerobic and aerobic conditions.
[0036] B. Identification of strains:
[0037] (1) In a sterile ultra-clean workbench, pick out a single colony and inoculate it in the corresponding liquid medium according to the shape, size, color, neatness of the edge, and whether the bulge is on th...
Embodiment 2
[0054] A preparation method for probiotics for infertility treatment, comprising the following steps:
[0055] 1) Isolate microorganisms from vaginal secretions, and cultivate several single colonies;
[0056] 2) carry out bacterial species identification to the microorganisms in the several single colonies, and obtain several bacterial strains;
[0057] 3) taking the several bacterial strains, co-incubating them with sperm cells in a liquid environment respectively, and screening out target bacterial strains based on the adhesion ability of each bacterial strain to sperm cells under microscope observation;
[0058] 4) Cultivate the target bacterial strain to prepare freeze-dried powder bacterial agent.
[0059] On the basis of the above technical solutions, the following conditions are met:
[0060] The medium used for isolating microorganisms in step 1) includes LB basal medium, MRS solid medium, BHI solid medium, BHI medium containing 5% (g / g) skimmed milk, and BSM solid ...
Embodiment 3
[0073] A preparation method for probiotics for infertility treatment, comprising the following steps:
[0074] 1) Isolate microorganisms from vaginal secretions, and cultivate several single colonies;
[0075] 2) carry out bacterial species identification to the microorganisms in the several single colonies, and obtain several bacterial strains;
[0076] 3) taking the several bacterial strains, co-incubating them with sperm cells in a liquid environment respectively, and screening out target bacterial strains based on the adhesion ability of each bacterial strain to sperm cells under microscope observation;
[0077] 4) Cultivate the target bacterial strain to prepare freeze-dried powder bacterial agent.
[0078] On the basis of the above technical solutions, the following conditions are met:
[0079] The medium used for isolating microorganisms in step 1) includes LB basal medium, MRS solid medium, BHI solid medium, BHI medium containing 5% (g / g) skimmed milk, and BSM solid ...
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