US2012088303A1PendingUtilityA1
Peptide fractions promoting growth and synthesis of desired product(s) into cell and/or tissue culture
Est. expiryDec 14, 2024(expired)· nominal 20-yr term from priority
C07K 14/57C12N 2500/76C12N 2500/92C12N 1/04C12P 21/06C12N 5/0018C12N 5/00C07K 14/415
25
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Claims
Abstract
The invention relates to preparing and/or supplementing a cell or tissue culture medium. In particular, said invention relates to a serum-free and/or protein-free cell culture medium comprising peptide fractions isolated from rapseeds, in particular rapseeds cakes. A method for the production of a cell culture comprising said peptide fractions and for the use thereof is also disclosed.
Claims
exact text as granted — not AI-modified1 . The use of a peptide extract for preparing and/or supplementing a serum-free in vitro cell or tissue culture medium, said extract being obtained by successive fractionations of a plant starting material, characterized in that said plant starting material consists of rapeseed.
2 . The use of a peptide extract for preparing and/or supplementing a protein-free in vitro cell or tissue culture medium, said extract being obtained by successive fractionations of a plant starting material, characterized in that said plant starting material consists of rapeseed.
3 . The use as claimed in claim 1 , characterized in that said peptide extract performs a function in terms of increasing the cell or tissue concentration and/or in terms of increasing the lifespan of the cells and/or in terms of the specific rate of production of one or more molecules of interest, said function being bound not solely by the elemental amino acid composition, but by the composition, of said amino acids, in peptide form.
4 . The use as claimed in claim 1 , characterized in that said plant starting material consists of rapeseed cake.
5 . The use as claimed in claim 1 , characterized in that said peptide extract consists of at least 80%, preferably at least 90%, by mass of nitrogenous material.
6 . The use as claimed in claim 5 , characterized in that said nitrogenous material consists of approximately 50% to approximately 60% of peptides having a molecular size of less than 500 daltons.
7 . The use as claimed in claim 5 , characterized in that said nitrogenous material consists of approximately 20% to approximately 30% of peptides having a molecular size of between 500 and 1000 daltons.
8 . The use as claimed in claim 1 , characterized in that said extract comprises between 20 and 40 mol % of aspartic acid and of glutamic acid and/or of their respective amides.
9 . The use as claimed in claim 1 , characterized in that said peptide extract comprises less than 1% by mass of phenolic compounds.
10 . The use as claimed in claim 1 , characterized in that said peptide extract has the following total amino acid composition:
Amino acids
Molar %
Alanine
2-6
Arginine
5-9
Aspartic acid + asparagine
10-14
Cysteine
0
Glutamic acid + Glutamine
15-19
Glycine
7-11
Histidine
1-5
Isoleucine
2-6
Leucine
5-9
Lysine
2-6
Méthionine
0-3
Phenylalanine
1-5
Proline
3-7
Serine
5-9
Threonine
5-9
Tryptophan
not
détermined
Tyrosine
1-5
Valine
5-9
11 . The use as claimed in claim 1 , for culturing cells, characterized in that said cells consist of eukaryotic cells, preferably animal cells.
12 . A serum-free cell or tissue culture medium obtained by using a peptide extract as claimed in claim 1 .
13 . The culture medium as claimed in claim 12 , characterized in that it also comprises vitamins, inorganic salts, free amino acids, organic acids and/or sugars.
14 . The use of a culture medium as claimed in claim 12 , for the bulk culturing of cells or tissues.
15 . A protein-free cell or tissue culture medium, obtained by using a peptide extract as claimed in claim 2 .
16 . The culture medium as claimed in claim 15 , characterized in that it also comprises vitamins, inorganic salts, free amino acids, organic acids and/or sugars.
17 . The use of a culture medium as claimed in claim 15 , for the bulk culturing of cells or tissues.
18 . A method of in vitro cell or tissue culture in a medium devoid of serum, characterized in that it consists in seeding said cells or tissues into a medium comprising a peptide extract obtained by successive fractionations of a plant starting material, said plant starting material consisting of rapeseed.
19 . The method as claimed in claim 18 , characterized in that said starting material consists of rapeseed cake.
20 . The method as claimed in claim 18 , characterized in that said extract consists of at least 80%, preferably at least 90%, by mass of nitrogenous material.
21 . The method as claimed in claim 18 , characterized in that said nitrogenous material consists of approximately 50% to approximately 60% of peptides having a molecular size of less than 500 daltons.
22 . The method as claimed in claim 18 , characterized in that said nitrogenous material consists of approximately 20% to approximately 30% of peptides having a molecular size of between 500 and 1000 daltons.
23 . The method as claimed in claim 18 , characterized in that said extract comprises between 20 and 40 mol % of aspartic acid and of glutamic acid and/or of their respective amides.
24 . The method as claimed in claim 18 , characterized in that said extract has the following total amino acid composition:
Amino acids
Molar %
Alanine
2-6
Arginine
5-9
Aspartic acid + asparagine
10-14
Cysteine
0
Glutamic acid + Glutamine
15-19
Glycine
7-11
Histidine
1-5
Isoleucine
2-6
Leucine
5-9
Lysine
2-6
Méthionine
0-3
Phenylalanine
1-5
Proline
3-7
Serine
5-9
Threonine
5-9
Tryptophan
not
determined
Tyrosine
1-5
Valine
5-9
25 . A method of in vitro cell or tissue culture in a protein-free medium, characterized in that it consists in seeding said cells or tissues into a medium comprising a peptide extract obtained by successive fractionations of a plant starting material, said plant starting material consisting of rapeseed.
26 . The method as claimed in claim 25 , characterized in that said starting material consists of rapeseed cake.
27 . The method as claimed in claim 25 , characterized in that said extract consists of at least 80%, preferably at least 90%, by mass of nitrogenous material.
28 . The method as claimed in claim 25 , characterized in that said nitrogenous material consists of approximately 50% to approximately 60% of peptides having a molecular size of less than 500 daltons.
29 . The method as claimed in claim 25 , characterized in that said nitrogenous material consists of approximately 20% to approximately 30% of peptides having a molecular size of between 500 and 1000 daltons.
30 . The method as claimed in claim 25 , characterized in that said extract comprises between 20 and 40 mol % of aspartic acid and of glutamic acid and/or of their respective amides.
31 . The method as claimed in claim 25 , characterized in that said extract has the following total amino acid composition:
Amino acids
Molar %
Alanine
2-6
Arginine
5-9
Aspartic acid + asparagine
10-14
Cysteine
0
Glutamic acid + Glutamine
15-19
Glycine
7-11
Histidine
1-5
Isoleucine
2-6
Leucine
5-9
Lysine
2-6
Méthionine
0-3
Phenylalanine
1-5
Proline
3-7
Serine
5-9
Threonine
5-9
Tryptophan
not
determined
Tyrosine
1-5
Valine
5-9
32 . A method for directly transferring an in vitro culture in a medium with serum, to a culture in a serum-free medium, characterized in that it consists in recovering the cells and/or the tissue from the medium with serum, and in seeding them or it directly into a medium as claimed in claim 12 .
33 . A method for adapting an in vitro culture in a medium with serum, to a culture in a serum-free medium, characterized in that it consists in recovering the cells and/or the tissue from the medium with serum, and in seeding them or it, stepwise, into a medium as claimed in claim 12 , said medium having decreasing serum concentrations, respectively.
34 . The method of adaptation as claimed in claim 33 , characterized in that said method of adaptation comprises the following four steps:
step 1: 75% medium with serum/25% serum-free medium+peptide extract, step 2: 50% medium with serum/50% serum-free medium+peptide extract, step 3: 25% medium with serum/75% serum-free medium+peptide extract, step 4: 100% serum-free medium+peptide extract.
35 . A method for directly transferring an in vitro culture in a medium with serum, to a culture in a protein-free medium, characterized in that it consists in recovering the cells and/or the tissue from the medium with serum, and in seeding them or it directly into a medium as claimed in claim 15 .
36 . A method for adapting an in vitro culture in a medium with serum, to a culture in a protein-free medium, characterized in that it consists in recovering the cells and/or the tissue from the medium with serum, and in seeding them or it, stepwise, into a medium as claimed in claim 15 , said medium having decreasing serum concentrations, respectively.
37 . The method of adaptation as claimed in claim 36 , characterized in that said method of adaptation comprises the following four steps:
step 1: 75% medium with serum/25% serum-free medium+peptide extract, step 2: 50% medium with serum/50% serum-free medium+peptide extract, step 3: 25% medium with serum/75% serum-free medium+peptide extract, step 4: 100% serum-free medium+peptide extract.
38 . A method for adapting an in vitro culture in a medium with serum, to a culture in a protein-free medium, characterized in that said method comprises the following steps:
step 1: carrying out the method as claimed in claim 32 , step 2: recovering the cells and/or the tissues from the serum-free medium, step 3: seeding said cells and/or tissues directly into a protein-free medium.
39 . A medium for culturing in vitro animal cells or tissues, which comprises an animal protein-free medium and a peptide extract obtained by successive fractionations of a plant starting material, wherein said plant starting material consists of rapeseed.
40 . The medium for culturing in vitro animal cells or tissues according to claim 39 , wherein said plant starting material consists of rapeseed cake.
41 . The medium for culturing in vitro animal cells or tissues according to claim 39 , wherein said extract consists of at least 80% by mass of nitrogenous material.
42 . The medium for culturing in vitro animal cells or tissues according to claim 39 , wherein said extract consists of at least 90% by mass of nitrogenous material.
43 . The medium for culturing in vitro animal cells or tissues according to claim 41 , wherein said nitrogenous material consists of approximately 50% to approximately 60% of peptides having a molecular size of less than 500 daltons.
44 . The medium for culturing in vitro animal cells or tissues according to claim 41 , wherein said nitrogenous material consists of approximately 20% to approximately 30% of peptides having a molecular size of between 500 and 1000 daltons.
45 . The medium for culturing in vitro animal cells or tissues according to claim 39 , wherein said extract comprises between 20 and 40 mol % of aspartic acid and of glutamic acid and/or of their respective amides.
46 . The medium for culturing in vitro animal cells or tissues according to claim 39 , wherein said extract has the following total amino acid composition:
Amino acids
Molar %
Alanine
2-6
Arginine
5-9
Aspartic acid + asparagine
10-14
Cysteine
0
Glutamic acid + Glutamine
15-19
Glycine
7-11
Histidine
1-5
Isoleucine
2-6
Leucine
5-9
Lysine
2-6
Méthionine
0-3
Phenylalanine
1-5
Proline
3-7
Serine
5-9
Threonine
5-9
Tryptophan
not
determined
Tyrosine
1-5
Valine
5-9Join the waitlist — get patent alerts
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