CN108315295A - The culture solution of testicular biopsy sperm - Google Patents

The culture solution of testicular biopsy sperm Download PDF

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Publication number
CN108315295A
CN108315295A CN201810262754.2A CN201810262754A CN108315295A CN 108315295 A CN108315295 A CN 108315295A CN 201810262754 A CN201810262754 A CN 201810262754A CN 108315295 A CN108315295 A CN 108315295A
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sperm
culture solution
raw material
pva
bsa
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许常龙
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Second Peoples Hospital of Nanning
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Second Peoples Hospital of Nanning
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0608—Germ cells
    • C12N5/061—Sperm cells, spermatogonia
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/12—Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/12—Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
    • C12N2500/14—Calcium; Ca chelators; Calcitonin
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/12—Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
    • C12N2500/16—Magnesium; Mg chelators
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/34—Sugars
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/50—Soluble polymers, e.g. polyethyleneglycol [PEG]

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Abstract

The present invention discloses a kind of culture solution of testicular biopsy sperm, is formulated by following raw material：NaCl 5.4~5.6 g/L, KCl 0.3~0.4 g/L, CaCl2﹒ 2H2O 0.2~0.4 g/L, KH2PO40.1~0.3g/L, MgSO4﹒ 7H2O 0.2~0.3 g/L, NaHCO30.3~0.4 4.5~5g/L of g/L, HEPES, 2.5~3 g/L of sodium lactate, 0.02~0.04 g/L of Sodium Pyruvate, 2~3mmol/L of pentoxifylline, 1~2 3~4 g/L of g/L, BSA of glucose, 0.05~0.08 g/L of penicillin, 0.02~0.06 g/L of streptomysin, phenol red 0.01~0.02 g/L, PVA 1~1.5 ‰, the present invention can effectively improve asthénospermie and asthenospermia disease, conducive to selecting for high-quality sperm, insemination quality is improved.

Description

The culture solution of testicular biopsy sperm
Technical field
The present invention relates to reproductive technology field, the culture solution of specifically a kind of testicular biopsy sperm.
Background technology
At present due to the influence of the various aspects factor such as environment, food, the infertile crowd of the mankind gradually increases, acceptor Work is inseminated or the people of test-tube baby is more and more, and there are many infertile reason, single that sexual function barrier is broadly divided into for male Azoospermia, oligospermatism, weak sperm can be further divided into normal two class of sexual function, the latter according to semen analysis result by hindering Disease, asthenospermia disease and sperm count normality infertility.
Artificial insemination or test-tube baby are typically that the sperm gone out with testicular biopsy and ovarian follicle slurry combine to obtain insemination ovum, For above-mentioned asthénospermie and asthenospermia disease, if directly sperm and the ovarian follicle slurry that testicular biopsy goes out are combined, gained The insemination archiblast amount arrived is bad, it is therefore necessary to overcome above-mentioned disadvantage, ensure to be possible to obtain in the case of sperm quality The insemination ovum of better quality, the present invention are to solve the problems, such as this.
Invention content
The technical problem to be solved by the present invention is to：A kind of culture solution of testicular biopsy sperm is provided, which has Effect improves asthénospermie and asthenospermia disease, conducive to selecting for high-quality sperm, improves insemination quality.
The technical solution adopted in the present invention is：A kind of culture solution of testicular biopsy sperm is provided, is prepared by following raw material It forms：NaCl 5.4~5.6 g/L, KCl 0.3~0.4 g/L, CaCl2﹒ 2H2O 0.2~0.4 g/L, KH2PO40.1~ 0.3g/L、MgSO4﹒ 7H2O 0.2~0.3 g/L, NaHCO30.3~0.4 4.5~5g/L of g/L, HEPES, sodium lactate 2.5~ 3 g/L, 0.02~0.04 g/L of Sodium Pyruvate, 2~3mmol/L of pentoxifylline, 1~2 3~4 g/ of g/L, BSA of glucose L, 0.05~0.08 g/L of penicillin, 0.02~0.06 g/L of streptomysin, phenol red 0.01~0.02 g/L, PVA 1~1.5 ‰.
Preferably, the culture solution is formulated by following raw material：NaCl 5.533 g/L、KCl 0.356 g/L、 CaCl2﹒ 2H2O 0.252 g/L、KH2PO4 0.162 g/L、MgSO4﹒ 7H2O 0.293 g/L、NaHCO3 0.349 g/L、 4.969 g/L of HEPES, 2.610 g/L of sodium lactate, 0.036 g/L of Sodium Pyruvate, pentoxifylline 2.50mmol/L, glucose 1.000 4.000 g/L of g/L, BSA, 0.060 g/L of penicillin, 0.050 g/L of streptomysin, phenol red 0.010 g/L, PVA 1‰。
The culture solution of testicular biopsy sperm of the present invention is used for the culture of testicular biopsy sperm, by that can improve weak essence after culture Sub- disease and asthenospermia disease, make motility of sperm greatly promote, and conducive to selecting for high-quality sperm, improve insemination quality.
Specific implementation mode
The present invention is described in detail below in conjunction with specific embodiment, the raw material in each embodiment is commercial product, HEPES is 4- hydroxyethyl piperazineethanesulfonic acids, and BSA is bovine serum albumin(BSA), and PVA is pharmaceutical grade Polyethylene alcohol.
Embodiment 1
The culture solution of the present embodiment is formulated by following raw material：NaCl 5.4 g/L、KCl 0.4 g/L、CaCl2﹒ 2H2O 0.2 g/L、KH2PO4 0.1g/L、MgSO4﹒ 7H2O 0.2 g/L、NaHCO30.4 g/L, HEPES 5g/L, 3 g/L of sodium lactate, Sodium Pyruvate 0.02g/L, pentoxifylline 2mmol/L, 13 g/L of g/L, BSA of glucose, 0.05 g/L of penicillin, streptomysin 0.02 g/L, phenol red 0.02 g/L, PVA 1 ‰, the content of PVA are thousand points of contents of raw material volume.
Embodiment 2
The culture solution of the present embodiment is formulated by following raw material：NaCl 5.6 g/L、KCl 0.3 g/L、CaCl2﹒ 2H2O 0.4 g/L、KH2PO4 0.1g/L、MgSO4﹒ 7H2O0.3 g/L、NaHCO30.3 g/L, HEPES 4.5g/L, sodium lactate 2.5g/ L, 0.04 g/L of Sodium Pyruvate, pentoxifylline 3mmol/L, 24 g/L of g/L, BSA of glucose, 0.08 g/L of penicillin, chain 0.06 g/L of mycin, phenol red 0.01 g/L, PVA1.5 ‰, the content of PVA are thousand points of contents of raw material volume.
Embodiment 3
The culture solution of the present embodiment is formulated by following raw material：NaCl 5.533 g/L、KCl 0.356 g/L、CaCl2﹒ 2H2O 0.252 g/L、KH2PO4 0.162 g/L、MgSO4﹒ 7H2O 0.293 g/L、NaHCO3 0.349 g/L、HEPES 4.969 g/L, 2.610 g/L of sodium lactate, 0.036 g/L of Sodium Pyruvate, pentoxifylline 2.50mmol/L, glucose 1.000 4.000 g/L of g/L, BSA, 0.060 g/L of penicillin, 0.050 g/L of streptomysin, phenol red 0.010 g/L, PVA 1 ‰, PVA's Content is thousand points of contents of raw material volume.
Embodiment 4
The culture solution of the present embodiment is formulated by following raw material：NaCl 5.523 g/L、KCl 0.335 g/L、CaCl2﹒ 2H2O 0.255 g/L、KH2PO4 0.152g/L、MgSO4﹒ 7H2O 0.253 g/L、NaHCO3 0.356 g/L、HEPES 4.586g/L, 2.558 g/L of sodium lactate, 0.028 g/L of Sodium Pyruvate, pentoxifylline 2.45mmol/L, 1.50 g/ of glucose L, BSA 3.56g/L, 0.068 g/L of penicillin, 0.045 g/L of streptomysin, phenol red 0.015 g/L, PVA 1.25 ‰.PVA's Content is thousand points of contents of raw material volume.
Embodiment 5
The culture solution of the present embodiment is formulated by following raw material：NaCl 5.485 g/L、KCl 0.388 g/L、CaCl2﹒ 2H2O 0.356 g/L、KH2PO4 0.25g/L、MgSO4﹒ 7H2O 0.258 g/L、NaHCO3 0.365 g/L、HEPES 4.668g/L, sodium lactate 2.65g/L, 0.035 g/L of Sodium Pyruvate, pentoxifylline 2.65mmol/L, 1.25 g/L of glucose, 3.55 g/L of BSA, 0.065 g/L of penicillin, 0.068 g/L of streptomysin, phenol red 0.015 g/L, PVA 1.35 ‰.PVA's contains Amount is thousand points of contents of raw material volume.
By the result data of the various embodiments described above and uses other common culture solutions in the prior art or trained without using The sperm of nutrient solution compares on vigor or activity respectively, and comparing result is as follows：
Embodiment Vigor（%） Motility rate（%）
Embodiment 1 10.2 16.5
Embodiment 2 9.8 15.4
Embodiment 3 14.8 20.1
Embodiment 4 10.4 15.8
Embodiment 5 11.3 16.4
Nostoc commune Vanch liquid 6.9 10.7

Claims (2)

1. a kind of culture solution of testicular biopsy sperm, it is characterised in that be formulated by following raw material：5.4~5.6 g/ of NaCl L, 0.3~0.4 g/L, CaCl of KCl2﹒ 2H2O 0.2~0.4 g/L, KH2PO40.1~0.3g/L, MgSO4﹒ 7H2O 0.2~ 0.3 g/L、NaHCO30.3~0.4 4.5~5g/L of g/L, HEPES, 2.5~3 g/L of sodium lactate, Sodium Pyruvate 0.02~ 0.04 g/L, 2~3mmol/L of pentoxifylline, 1~2 3~4 g/L of g/L, BSA of glucose, 0.05~0.08 g/ of penicillin L, 0.02~0.06 g/L of streptomysin, phenol red 0.01~0.02 g/L, PVA 1~1.5 ‰.
2. the culture solution of testicular biopsy sperm according to claim 1, it is characterised in that be formulated by following raw material： NaCl 5.533 g/L、KCl 0.356 g/L、CaCl2﹒ 2H2O 0.252 g/L、KH2PO4 0.162 g/L、MgSO4﹒ 7H2O 0.293 g/L、NaHCO30.349 4.969 g/L of g/L, HEPES, 2.610 g/L of sodium lactate, 0.036 g/L of Sodium Pyruvate, Pentoxifylline 2.50mmol/L, 1.000 4.000 g/L of g/L, BSA of glucose, 0.060 g/L of penicillin, streptomysin 0.050 g/L, phenol red 0.010 g/L, PVA 1 ‰.
CN201810262754.2A 2018-03-28 2018-03-28 The culture solution of testicular biopsy sperm Pending CN108315295A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104046590A (en) * 2014-06-18 2014-09-17 郜鸿生物科技（上海）有限公司 In-vitro culture medium for sperm from oligospermia and asthenospermia patients
CN107034179A (en) * 2017-05-25 2017-08-11 四川大学华西第二医院 A kind of solution for improving the detection of Human Testis sperm

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104046590A (en) * 2014-06-18 2014-09-17 郜鸿生物科技（上海）有限公司 In-vitro culture medium for sperm from oligospermia and asthenospermia patients
CN107034179A (en) * 2017-05-25 2017-08-11 四川大学华西第二医院 A kind of solution for improving the detection of Human Testis sperm

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