WO2008016958A2 - Compositions de milieux de cellules reproductrices et procédés destinés à améliorer la fertilité - Google Patents

Compositions de milieux de cellules reproductrices et procédés destinés à améliorer la fertilité Download PDF

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Publication number
WO2008016958A2
WO2008016958A2 PCT/US2007/074935 US2007074935W WO2008016958A2 WO 2008016958 A2 WO2008016958 A2 WO 2008016958A2 US 2007074935 W US2007074935 W US 2007074935W WO 2008016958 A2 WO2008016958 A2 WO 2008016958A2
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WO
WIPO (PCT)
Prior art keywords
composition
derived
colostrum
reproductive
cell
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Application number
PCT/US2007/074935
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English (en)
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WO2008016958B1 (fr
WO2008016958A3 (fr
Inventor
Ludwig Simmet
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Minitube Of America, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Application filed by Minitube Of America, Inc. filed Critical Minitube Of America, Inc.
Publication of WO2008016958A2 publication Critical patent/WO2008016958A2/fr
Publication of WO2008016958A3 publication Critical patent/WO2008016958A3/fr
Publication of WO2008016958B1 publication Critical patent/WO2008016958B1/fr

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Classifications

    • 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
    • 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
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0604—Whole embryos; Culture medium therefor
    • 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
    • C12N2500/70—Undefined extracts
    • C12N2500/80—Undefined extracts from animals
    • C12N2500/84—Undefined extracts from animals from mammals
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/105—Insulin-like growth factors [IGF]
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/155—Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
    • 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
    • C12N2517/00—Cells related to new breeds of animals
    • C12N2517/10—Conditioning of cells for in vitro fecondation or nuclear transfer

Definitions

  • Bovine Spongiform Encephalopathy is a transmissible, neurodegenerative disorder in cattle that was first recognized in the United Kingdom in 1986. Subsequently, the discovery of an indigenous, North American case of BSE in Canada in 2003 provoked international concern about safe sourcing of bovine materials. Non-affected countries prohibited the importation and use of materials derived from cattle in food and cosmetic products, resulting in a disruption of international trade in bovine products. Some of these bans have since been partially lifted. However, many countries continue to prohibit the importation of bovine meat and blood products from BSE-affected countries.
  • Bovine serum has been used to prepare a number of biological products, including cell culture products.
  • BSA bovine serum albumin
  • FDA Food and Drug Administration
  • EU European Union
  • the invention provides a composition comprising a reproductive cell medium and a milk- or colostrum-derived growth factor.
  • the invention provides a composition comprising a reproductive cell medium, ⁇ -lactalbumin and a growth factor.
  • the invention provides a method of increasing the number of offspring of an artificially inseminated mammal, comprising inseminating the mammal with semen and a composition comprising a reproductive cell medium comprising a milk- or colostrum-derived growth factor.
  • the invention provides a method of increasing the number of offspring of an artificially inseminated mammal, comprising inseminating the mammal with semen and a composition comprising a reproductive cell medium, ⁇ - lactalbumin and a growth factor.
  • the invention is related to colostrum- and milk-derived biosubstitutes for blood and serum products in general, and for use in cell media compositions in particular. Not only does the present invention circumvent restrictions on importation and use of bovine serum products, cost advantages may also be realized by use of the present invention.
  • the inventors have discovered that replacement of certain serum products with colostrum- and milk-derived equivalents in sperm culture medium surprisingly results in significantly improved fertilization rates in some in vitro fertilization protocols, although no particular degree of improvement in fertilization rates are required for the benefits of the present compositions to be realized.
  • the invention provides colostrum- and milk-derived products for use in cell media compositions.
  • the cell media compositions are reproductive cell media compositions, wherein the reproductive cells to be cultured are mammalian reproductive cells. More suitably, the cell media compositions are sperm cell media compositions wherein the sperm cells to be cultured are mammalian sperm cells.
  • reproductive cells encompasses sperm cells, oocytes, and embryos of any mammal, bird, or fish, including livestock (e.g., pigs, cows, horses, sheep and the like) and humans.
  • a "reproductive cell medium” is a medium used for the collection, holding, processing, in vitro fertilization, sexing, culturing, or storing (including long-term cryopreservation) of mammalian, avian, or piscian reproductive cells, and includes both solid and liquid compositions, as well as solid compositions that are reconstituted or mixed with a liquid carrier, such as water, for use.
  • a liquid carrier such as water
  • the reproductive cell medium is formulated to maintain the viability of the reproductive cells.
  • sperm cell medium refers to any medium used for the collection, holding, processing, in vitro fertilization, sexing, culturing, or storing (including long-term cryopreservation) of mammalian, avian or piscian sperm cells and/or semen.
  • the sperm cell medium may serve to maintain the viability of the sperm cells and to increase the volume of semen.
  • Milk- and colostrum-derived products suitable for use in the present invention are commercially available, or may be purified from milk or colostrum using standard methods.
  • An example of a commercially available milk-derived product is ⁇ - lactalbumin from BIOPURE Corp.
  • a "milk-derived” or “colostrum-derived” product is any biological product purified from milk or colostrum, respectively, using standard purification methods.
  • the term "purified” refers to material that is at least partially separated from components which normally accompany it in its native state. Purity of polypeptides is typically determined using analytical techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography.
  • a polypeptide that is the predominant species present in a preparation is “substantially purified.”
  • purified denotes that a preparation containing the polypeptide may give rise to essentially one band in an electrophoretic gel.
  • polypeptide refers to two or more amino acid moieties linked by amide bonds, and includes peptides and proteins.
  • substantially homology refers to a polypeptide sequence that performs substantially the same function as the parent sequence and has at least 60%, or more preferably, 75%, and most preferably, 90% or 95% or greater, sequence identity.
  • suitable colostrum-derived products are growth factors.
  • the growth factors are IGF-1 and/or TGF- ⁇ 2.
  • colostrum-derived IGF-1 is included in sperm cell media compositions at a concentration ranging from about 50 ng/L to about 500 ng/L. More suitably, IGF-1 is included in sperm cell media compositions at a concentration ranging from about 100 ng/L to about 200 ng/L.
  • colostrum-derived IGF-1 is included at a concentration of about 160 ng/L
  • colostrum-derived TGF- ⁇ 2 is included in sperm cell media compositions at a concentration ranging from about 10 ng/L to about 200 ng/L. More suitably, colostrum-derived TGF- ⁇ 2 is included in sperm cell media compositions at a concentration ranging from about 40 ng/L to about 100 ng/L. Most suitably, colostrum-derived TGF- ⁇ 2 is included at a concentration of about 55 ng/L.
  • Cell media compositions suitably comprise both IGF-1 and TGF- ⁇ 2.
  • a suitable ratio of IGF-1 to TGF ⁇ 2 is about 5:1 to about 2:1. Most suitably, the ratio of IGF-1 to TGF ⁇ 2 is about 3:1.
  • a suitable milk-derived product is ⁇ -lactalbumin.
  • ⁇ -lactalbumin is included in reproductive cell media compositions at concentrations ranging from about 1.0 to about 5.0 g/L. Most suitably, ⁇ -lactalbumin is included in reproductive cell media compositions at a concentration of about 2.5 g/L.
  • the composition comprises cell media, colostrum-derived IGF-1 , colostrum-derived TGF- ⁇ 2 and milk- derived ⁇ -lactalbumin.
  • the composition also suitably includes a reproductive cell, e.g., a sperm cell or oocyte. Most suitably, the composition includes a porcine or bovine sperm cell or oocyte.
  • compositions of the invention are substantially free of bovine serum-derived products.
  • a "serum-derived product” is any biological product purified from serum using standard purification methods.
  • Another embodiment of the invention provides a method of increasing the number of offspring of an artificially inseminated mammal, comprising inseminating the mammal with a composition comprising sperm and a milk- and/or colostrum-derived product.
  • the number of offspring is increased relative to a suitable control.
  • a suitable control is a mammal artificially inseminated with a composition comprising sperm, but not comprising a milk- or colostrum-derived product.
  • An additional suitable control is a mammal artificially inseminated with a composition comprising sperm and a bovine serum-derived product, but not comprising a milk- or colostrum-derived product.
  • the artificially inseminated mammal is a pig or a cow.
  • a sperm cell culture media was formulated to include the following ingredients:
  • Example 2 Fetal recovery using media formulated with serum-derived versus colostrum- derived additives.
  • Gilts were inseminated with one of three compositions or a control composition, as defined below, and were killed after the first trimester.
  • the artificial insemination (Al) compositions were as follows:
  • X1 ENDURAGUARD (Minitube, Verona, Wl) (with 2.5 g/L BSA and serum-derived TGF- ⁇ 2 (9.3 ng/L) and serum-derived IGF-1 (450 ng/L)) + sperm cells (500 x 10 6 ).
  • X2 ENDURAGUARD (Minitube, Verona, Wl) base formulation (with 2.5 g/L BSA and colostrum-derived IGF-1 (160 ng/L) and colostrum-derived TGF ⁇ 2 (55 ng/L)) + sperm cells (500 x 10 6 ).
  • X3 ENDURAGUARD (Minitube, Verona, Wl) base formulation (with 2.5 g/L ⁇ -lactalbumin and colostrum-derived IGF-1 (160 ng/L) and TGF ⁇ 2 (55 ng/L)) + sperm cells (500 x 10 6 ).
  • the X3 composition containing ⁇ -lactalbumin, colostrum- derived IGF-1 and colostrum-derived TGF ⁇ 2 resulted in a trend suggesting improvement in the average number of fetuses per gilt in comparison to the X1 and X2 compositions.
  • Example 3 Fetal recovery using media formulated with serum-derived versus colostrum- derived additives.
  • Gilts are inseminated with compositions described below, and are killed after the first trimester.
  • X2 ENDURAGUARD (Minitube, Verona, Wl) (with 2.5 g/ L ⁇ -lactalbumin and serum-derived TGF- ⁇ 2 (9.3 ng/L) and IGF-1 (450 ng/L)) + sperm cells (500 x 106).
  • X2 composition is expected to result in an improvement in the average number of fetuses per gilt in comparison to the X1 composition.
  • any numerical value recited herein includes all values from the lower value to the upper value, i.e., all possible combinations of numerical values between the lowest value and the highest value enumerated are to be considered to be expressly stated in this application. For example, if a concentration range is stated as 1% to 50%, it is intended that values such as 2% to 40%, 10% to 30%, or 1% to 3%, etc., are expressly enumerated in this specification. These are only examples of what is specifically intended.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Reproductive Health (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Gynecology & Obstetrics (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La présente invention concerne des compositions pour cellules reproductrices qui contiennent un milieu de cellule reproductrive approprié et un produit dérivé du lait ou du colostrum. Ce produti dérivé du lait ou du colostrum peut contenir un facteur de croissance tel qu'un facteur de croissance transformant ou un facteur de croissance de type insuline ou encore α-lactalbumin. Les compositions peuvent éventuellement être dépourvues de produits dérivés de sérum de bovin. L'invention concerne également les procédés utilisant ces compositions destinés à améliorer la fertilité des animaux.
PCT/US2007/074935 2006-08-01 2007-08-01 Compositions de milieux de cellules reproductrices et procédés destinés à améliorer la fertilité WO2008016958A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US82104406P 2006-08-01 2006-08-01
US60/821,044 2006-08-01

Publications (3)

Publication Number Publication Date
WO2008016958A2 true WO2008016958A2 (fr) 2008-02-07
WO2008016958A3 WO2008016958A3 (fr) 2008-03-20
WO2008016958B1 WO2008016958B1 (fr) 2008-05-08

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ITRM20100300A1 (it) * 2010-06-03 2011-12-04 Lo Li Pharma Srl Formulazione di composto per il trattamento del liquido seminale maschile destinato alla tecniche di riproduzione medicalmente assistita
WO2015095650A1 (fr) * 2013-12-19 2015-06-25 Puretein Bioscience Llc. Méthodes de traitement d'un animal

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4511558A (en) * 1983-05-16 1985-04-16 University Patents, Inc. Alpha-lactalbumin contraceptive
US20030157473A1 (en) * 2002-02-21 2003-08-21 Minitube Of America Compositions comprising reproductive cell media and methods for using such compositions

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4511558A (en) * 1983-05-16 1985-04-16 University Patents, Inc. Alpha-lactalbumin contraceptive
US20030157473A1 (en) * 2002-02-21 2003-08-21 Minitube Of America Compositions comprising reproductive cell media and methods for using such compositions
US20040076945A1 (en) * 2002-02-21 2004-04-22 Minitube Of America, Inc. Compositions comprising reproductive cell media and methods for using such compositions
US6849394B2 (en) * 2002-02-21 2005-02-01 Minitube Of America Compositions comprising reproductive cell media and methods for using such compositions

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ITRM20100300A1 (it) * 2010-06-03 2011-12-04 Lo Li Pharma Srl Formulazione di composto per il trattamento del liquido seminale maschile destinato alla tecniche di riproduzione medicalmente assistita
WO2015095650A1 (fr) * 2013-12-19 2015-06-25 Puretein Bioscience Llc. Méthodes de traitement d'un animal
US10279013B2 (en) 2013-12-19 2019-05-07 Puretein Bioscience Llc Methods for treating an animal

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Publication number Publication date
WO2008016958B1 (fr) 2008-05-08
WO2008016958A3 (fr) 2008-03-20

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