CA1165707A - Method of tissue culture - Google Patents
Method of tissue cultureInfo
- Publication number
- CA1165707A CA1165707A CA000386671A CA386671A CA1165707A CA 1165707 A CA1165707 A CA 1165707A CA 000386671 A CA000386671 A CA 000386671A CA 386671 A CA386671 A CA 386671A CA 1165707 A CA1165707 A CA 1165707A
- Authority
- CA
- Canada
- Prior art keywords
- extract
- culture medium
- algae
- cells
- animal
- Prior art date
- Legal status (The legal status 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 status listed.)
- Expired
Links
Abstract
Abstract of the Disclosure In a tissue culture of animal cells, it is made possible to perform successive cultivation of animal tissue efficiently under sparing or even without use of animal serum which is indispensable in the conventional method, by conducting the tissue culture using a culture medium containing an extract of micro algae such as Chlorella, Scenedesmus, Spirulina and so on.
Description
11~ 7~i7 Title of the Invention . .
METHOD OF TISSUE CULTURE
Back~round of the Invention 1. Field of the Invention The present invention relates to a method of tissue culture of animal tissue and more particularly to a method of tissue culture which enables an efficient successive cultivation of somatic cells of animal~to be maintained.
METHOD OF TISSUE CULTURE
Back~round of the Invention 1. Field of the Invention The present invention relates to a method of tissue culture of animal tissue and more particularly to a method of tissue culture which enables an efficient successive cultivation of somatic cells of animal~to be maintained.
2. Description of the Prior Art It has heretofore been necessary to employ so-called successive cultivation of animal tissue for wide categories of biological studies including fields such as medicine, pharmacology and so on as an effective and practicable means of research.
For this reason, the successive cultivation of animal tissue has found its application in various fields of research, such as in elucidating the mechanisms of cell differentiation, cancer etc. in recent years.
For successive cultivation of animal tissue, it is necesqary to use, excluding a few exceptions, a serum of, such as, adult, neonate or fetus of cattle, horse, chicken, rabbit and so on, in addition to the chemically defined synthetic culture medium consisting of amino acids, vitamins, minerals and so on. The reason for the indispensability of serum is based on the fact, that a causal matter of cell multiplicative growth factor ~a substance other than the general nutritive materials) is contained therein, so that a multiplication of animal tissue will be impossi~le without serum, owing to the lack of the said :,, ,, ~
causal matter. There are yet many unknown aspects besides the problem of instability of the matter. In addition, serum is expen-sive due to the insufficient supply system therefore. For these reasons, a tissue culture has remained as difficult.
S BRIEF SUMMARY OF THE INVENTION
Thus this invention is concerned with a method of tissue culture which permits the resolution of difficulties described above.
It is hoped to provide a method of tissue culture in which a whole or a part of the animal serum to be incorporated in the culture medium can be dispensed with.
It is also of concern to provide a method of tissue culture which enables successive cultivation in an efficient manner.
The invention will appear more clearly from the following description with the accompanying drawings.
The method according to the present invention is charac-terized in cultivating animal tissue under the employment of a culture medium containing an extract of micro algae.
More particularly, the invention provides a method for culturing cells or tissue of lower animals comprising culturing a sample of cells or tissue of a lower animal in a nutritive culture medium for cells and tissues of lower animals, said culture medium containing an effective amount of an extract of algae capable of causing cell multiplication of said cells or tissue.
In a more preferred aspect, the invention provides a method for culturing tissues and cells of lower animals, which comprises:
; providing a nutrient culture medium containing, per ml of said -~L
1~65707 culture medium, from 0.3 to 400~/g, calculated as the solids, of an aqueous extract of an algae selected from the group consisting of Chlorella, Scenedesmus, and Spirulina, said extract having been prepared by extracting said algae with water at a temperature of from 50C to 150C, for from 0.5 to 120 minutes, so that said extract contains sugars, proteins, polysaccharides and nucleic acids having molecular weights in the range of from 1,000 to 1,000,000 and exhibits an activity for promoting multiplication of animal cells; then placing and culturing on said culture medium a sample which is a piece of animal tissue or animal cells obtained from a lower animal, which sample is free of human cells and human tissues.
BRIEF DESCRIPTION OF THE DRAWINGS
Fig. 1 is a chart of molecular fractionation of Al-component of hot water extract of Chlorella cells.
Figs. 2 and 3 show graphically the results obtained in the experiments given in Examples 1 and 2 respectively.
A
., ., ~.,~ , -` 1165707 _ tailed Description of the Invention In this specification, the term "micro algae" means unicellular algae or its near relatives, such as for example, Chlorella, Scenedesmus, Spirulina or so on inclusive of natural living and cultivated ones.
The extract of micro algae as indicated in this speci-fication denotes an extract which is from an extraction of cells of one or more of said micro algae using an adequate solvent. As the solvent, an aqueous one is especially preferred.
Among the aqueous solvents, such as for example, water itself and aqueous solutions containing dissolved therein acids, bases and organic solvent~, may be exemplified.
In order to effect the extraction, algae cells are brought into contact with the solvent which is heated or kept at ordinary temperature. It is preferable to employ a hot water extraction, in which the algae cells are suspended in water in an amount of 1 - 1000 grams in dry weight of algae per liter of water and are ~ept at 50 - lS0 C for 0.5 - 120 minutes, preferably at 100 C for more than 1 minute, and then are removed from water by, for example, centrifugation etc., to leave an extract.
The extract may be refined on requirement by means of, for example, gel filtration, dialysis and so on.
The thus obtained extract of micro algae contains sugars, proteins, polysaccharides, nucleic acid and 80 on having molecular weight~ in the range from 1,000 to 1,000,000, and exhibits a multipllcation promoting activity in cultivating animal tissue.
The extract obtained as above can be used per se, as a fraction of molecular weight fractionation thereof or in a form
For this reason, the successive cultivation of animal tissue has found its application in various fields of research, such as in elucidating the mechanisms of cell differentiation, cancer etc. in recent years.
For successive cultivation of animal tissue, it is necesqary to use, excluding a few exceptions, a serum of, such as, adult, neonate or fetus of cattle, horse, chicken, rabbit and so on, in addition to the chemically defined synthetic culture medium consisting of amino acids, vitamins, minerals and so on. The reason for the indispensability of serum is based on the fact, that a causal matter of cell multiplicative growth factor ~a substance other than the general nutritive materials) is contained therein, so that a multiplication of animal tissue will be impossi~le without serum, owing to the lack of the said :,, ,, ~
causal matter. There are yet many unknown aspects besides the problem of instability of the matter. In addition, serum is expen-sive due to the insufficient supply system therefore. For these reasons, a tissue culture has remained as difficult.
S BRIEF SUMMARY OF THE INVENTION
Thus this invention is concerned with a method of tissue culture which permits the resolution of difficulties described above.
It is hoped to provide a method of tissue culture in which a whole or a part of the animal serum to be incorporated in the culture medium can be dispensed with.
It is also of concern to provide a method of tissue culture which enables successive cultivation in an efficient manner.
The invention will appear more clearly from the following description with the accompanying drawings.
The method according to the present invention is charac-terized in cultivating animal tissue under the employment of a culture medium containing an extract of micro algae.
More particularly, the invention provides a method for culturing cells or tissue of lower animals comprising culturing a sample of cells or tissue of a lower animal in a nutritive culture medium for cells and tissues of lower animals, said culture medium containing an effective amount of an extract of algae capable of causing cell multiplication of said cells or tissue.
In a more preferred aspect, the invention provides a method for culturing tissues and cells of lower animals, which comprises:
; providing a nutrient culture medium containing, per ml of said -~L
1~65707 culture medium, from 0.3 to 400~/g, calculated as the solids, of an aqueous extract of an algae selected from the group consisting of Chlorella, Scenedesmus, and Spirulina, said extract having been prepared by extracting said algae with water at a temperature of from 50C to 150C, for from 0.5 to 120 minutes, so that said extract contains sugars, proteins, polysaccharides and nucleic acids having molecular weights in the range of from 1,000 to 1,000,000 and exhibits an activity for promoting multiplication of animal cells; then placing and culturing on said culture medium a sample which is a piece of animal tissue or animal cells obtained from a lower animal, which sample is free of human cells and human tissues.
BRIEF DESCRIPTION OF THE DRAWINGS
Fig. 1 is a chart of molecular fractionation of Al-component of hot water extract of Chlorella cells.
Figs. 2 and 3 show graphically the results obtained in the experiments given in Examples 1 and 2 respectively.
A
., ., ~.,~ , -` 1165707 _ tailed Description of the Invention In this specification, the term "micro algae" means unicellular algae or its near relatives, such as for example, Chlorella, Scenedesmus, Spirulina or so on inclusive of natural living and cultivated ones.
The extract of micro algae as indicated in this speci-fication denotes an extract which is from an extraction of cells of one or more of said micro algae using an adequate solvent. As the solvent, an aqueous one is especially preferred.
Among the aqueous solvents, such as for example, water itself and aqueous solutions containing dissolved therein acids, bases and organic solvent~, may be exemplified.
In order to effect the extraction, algae cells are brought into contact with the solvent which is heated or kept at ordinary temperature. It is preferable to employ a hot water extraction, in which the algae cells are suspended in water in an amount of 1 - 1000 grams in dry weight of algae per liter of water and are ~ept at 50 - lS0 C for 0.5 - 120 minutes, preferably at 100 C for more than 1 minute, and then are removed from water by, for example, centrifugation etc., to leave an extract.
The extract may be refined on requirement by means of, for example, gel filtration, dialysis and so on.
The thus obtained extract of micro algae contains sugars, proteins, polysaccharides, nucleic acid and 80 on having molecular weight~ in the range from 1,000 to 1,000,000, and exhibits a multipllcation promoting activity in cultivating animal tissue.
The extract obtained as above can be used per se, as a fraction of molecular weight fractionation thereof or in a form
- 3 -'.~''1~
11657~7 of concentrate or dry powder obtained by freeze-drying, spray-drying etc. Especially, a high molecular fraction containing sugars, proteins and polysaccharides and dry powder thereof is preferred.
According to the present invention, animal tissue is cultivated using a culture medium containing an extract of micro algae, wherein the procedures of culture, basic culture medium and so on may be identical with those in the conventional tissue culture. Thus, a known synthetic culture medium contain-ing amino acids, vitamins and so on can be used as the basic culture medium, to which an extract mentioned above is added to perform the cultivation of animal tissue under aseptic con-ditions.
The animal tissue to be cultured includes somatic cells taken out of individual animal body, such as normal tissue and cancer cells.
The amount of addition of the micro algae extract to the basic culture medium may be in the range from 0.3 to 400 ~g and preferably in the range from 1 to 100 ~g in dry weight of the extracted matter per ml of the medium. The culture medium to which the micro algae extract has been added can be used per se for the cultivation, while it is also possible to incor-porate a serum addition as in the culture medium of prior art téchniques. Here, it is possible to reduce the amount of addi-tion of serum, which has heretofore been assumed to amount to about 10 %, up to a value of 1 %.
When animaltissues are cultivated by a culture medium containing the micro algae extract, the successive cultivation 7'~
" ' ' ~ .
, ' .
`~ 116S7~17 can be maintained by the multiplication promotion effect.
Comparable effects can be achieved, if the amount of serum which heretofore is considered in general as necessary to add in an amount of lO % is reduced to l/lO, so that a great reduction of serum amount can be reached.
Yet the cause for rendering the successive cultivation of animal tissue possible is not clear. It is to be assumed that the metabolism of the tissue is stimulated and becomes prosperous by the action of an unknown biologically active substance contained in the high molecular fraction of the micro algae extract, which contains especially glycoprotein, poly-saccharide and so on.
As described above in detail, the present invention brings forth substantial advantages, that the culture of animal tissue can be performed without employment or, even in case of employment, with only a little amount of addition of animal ~erum which i8 uhstable not only in the composition but also in the supply thereof and is disadvantageous from an economical point of view and that the tissue culture can be carried out ln An efficient and stable manner.
Description of the Preferred Embodiment The present invention is further explained by the following Examples.
Exam ~ 1 30 g of Chlorella powder were suspended in l Q of water and were ~ubjected to hot water extraction at lO0 C for 30 minute8. The suspen~lon was then centrifuged and the ''v , . ., ' ~
.
: '' ' '~ :
supernatant (corresponding to a dry extracted matter of 4.5 g) was caused to undergo a molecular fractionation on Sephadex G-25 column (trade mark of the firm Phamacia). The fractions with molecular weight~ above 3,000 were treated by adsorbing on DEAE-cellulose column (a trade mar~ of the firm Brown).
The so adsorbed column was then eluted by a M/100 carbonate buffer. A fraction having a component with molecular weight over 70,000, and in the range from 30,000 to 10,000 which contains rich neutral sugar and a component with molecular weight in the range from 10,000 to 3,000 which contains neutral sugar and protein in nearly equivalent proportion was obtained ~which i5 denoted hereinafter as Fraction Al) and this Fraction Al was dried by freezing to prepare 0.63 g of dry matter. The molecular fractionation chart of Fraction Al is given in Fig. 1.
S X 104 of cell strain RLC-10 (rat liver cells) were cultivated in a culture medium prepared by adding a fetus cattle ~erum (FCS) in an amount of 10 ~ to the basic culture medium DM-160 indicated in Table 1 ~Sector C). In a similar manner, the said cell strain RLC-10 were cultivated in a culture medium prepared by adding said FCS in an amount of 1 ~ and the above mentioned Chlorella Fraction A~ in an amount of 5 ~g/ml to the basic culture medium DM-160 (Sector T). The multiplication of the cell~ were examined by comparing the above two cultivation sectors. Both sector~ showed a similar trend of multiplication of cells. The results obtained are shown graphically in Fig. 2.
In a ~imilar manner a~ in Example 1, cell strain JTC-15 (rat ascite~ hepatoma), JTC-2 (rat ascites hepatoma), RLG-l ., - ` ' ' '......................... '' ' ' .
1~657~7 (rat lung cells) and RLC-18 ~rat liver cells) were examined for their multiplication rates in the medium as employed in Example 1 with variable contents, i.e. 0.5 ~g/ml, 5 ~g/ml and 50 ~g/ml, of Chlorella Fraction Al. The tendencies of the multiplication of the cells were the same. The results obtained are shown graphically in Fig. 3.
:~7 Table 1 Composition of Basic Culture Medium DM-160 Amino Acidsmg/l Vi tamins mg/l . .
Ala 400 B~ 1.0 Arg 100 B2 1. 0 A5p 25 B 6 1 . O
Asn 25 B 12 0 . 005 Cys-hCl 80 Pantothenic 1.0 acid Glu 150 Nicotinamide 1.0 Gln 300 Biotin 0.1 Gly 15 Choline HCl 5 . 0 His 30 C 1.0 Ile 150 Folic acid 1.0 Leu 400 Inositol 5.0 Lys 100 Met 80 Phe 80 Pro 12 Ser 8 0 Thr 100 ~rp 40 Tyr 5 0 Val 85 ,~ .
:
11657~7 of concentrate or dry powder obtained by freeze-drying, spray-drying etc. Especially, a high molecular fraction containing sugars, proteins and polysaccharides and dry powder thereof is preferred.
According to the present invention, animal tissue is cultivated using a culture medium containing an extract of micro algae, wherein the procedures of culture, basic culture medium and so on may be identical with those in the conventional tissue culture. Thus, a known synthetic culture medium contain-ing amino acids, vitamins and so on can be used as the basic culture medium, to which an extract mentioned above is added to perform the cultivation of animal tissue under aseptic con-ditions.
The animal tissue to be cultured includes somatic cells taken out of individual animal body, such as normal tissue and cancer cells.
The amount of addition of the micro algae extract to the basic culture medium may be in the range from 0.3 to 400 ~g and preferably in the range from 1 to 100 ~g in dry weight of the extracted matter per ml of the medium. The culture medium to which the micro algae extract has been added can be used per se for the cultivation, while it is also possible to incor-porate a serum addition as in the culture medium of prior art téchniques. Here, it is possible to reduce the amount of addi-tion of serum, which has heretofore been assumed to amount to about 10 %, up to a value of 1 %.
When animaltissues are cultivated by a culture medium containing the micro algae extract, the successive cultivation 7'~
" ' ' ~ .
, ' .
`~ 116S7~17 can be maintained by the multiplication promotion effect.
Comparable effects can be achieved, if the amount of serum which heretofore is considered in general as necessary to add in an amount of lO % is reduced to l/lO, so that a great reduction of serum amount can be reached.
Yet the cause for rendering the successive cultivation of animal tissue possible is not clear. It is to be assumed that the metabolism of the tissue is stimulated and becomes prosperous by the action of an unknown biologically active substance contained in the high molecular fraction of the micro algae extract, which contains especially glycoprotein, poly-saccharide and so on.
As described above in detail, the present invention brings forth substantial advantages, that the culture of animal tissue can be performed without employment or, even in case of employment, with only a little amount of addition of animal ~erum which i8 uhstable not only in the composition but also in the supply thereof and is disadvantageous from an economical point of view and that the tissue culture can be carried out ln An efficient and stable manner.
Description of the Preferred Embodiment The present invention is further explained by the following Examples.
Exam ~ 1 30 g of Chlorella powder were suspended in l Q of water and were ~ubjected to hot water extraction at lO0 C for 30 minute8. The suspen~lon was then centrifuged and the ''v , . ., ' ~
.
: '' ' '~ :
supernatant (corresponding to a dry extracted matter of 4.5 g) was caused to undergo a molecular fractionation on Sephadex G-25 column (trade mark of the firm Phamacia). The fractions with molecular weight~ above 3,000 were treated by adsorbing on DEAE-cellulose column (a trade mar~ of the firm Brown).
The so adsorbed column was then eluted by a M/100 carbonate buffer. A fraction having a component with molecular weight over 70,000, and in the range from 30,000 to 10,000 which contains rich neutral sugar and a component with molecular weight in the range from 10,000 to 3,000 which contains neutral sugar and protein in nearly equivalent proportion was obtained ~which i5 denoted hereinafter as Fraction Al) and this Fraction Al was dried by freezing to prepare 0.63 g of dry matter. The molecular fractionation chart of Fraction Al is given in Fig. 1.
S X 104 of cell strain RLC-10 (rat liver cells) were cultivated in a culture medium prepared by adding a fetus cattle ~erum (FCS) in an amount of 10 ~ to the basic culture medium DM-160 indicated in Table 1 ~Sector C). In a similar manner, the said cell strain RLC-10 were cultivated in a culture medium prepared by adding said FCS in an amount of 1 ~ and the above mentioned Chlorella Fraction A~ in an amount of 5 ~g/ml to the basic culture medium DM-160 (Sector T). The multiplication of the cell~ were examined by comparing the above two cultivation sectors. Both sector~ showed a similar trend of multiplication of cells. The results obtained are shown graphically in Fig. 2.
In a ~imilar manner a~ in Example 1, cell strain JTC-15 (rat ascite~ hepatoma), JTC-2 (rat ascites hepatoma), RLG-l ., - ` ' ' '......................... '' ' ' .
1~657~7 (rat lung cells) and RLC-18 ~rat liver cells) were examined for their multiplication rates in the medium as employed in Example 1 with variable contents, i.e. 0.5 ~g/ml, 5 ~g/ml and 50 ~g/ml, of Chlorella Fraction Al. The tendencies of the multiplication of the cells were the same. The results obtained are shown graphically in Fig. 3.
:~7 Table 1 Composition of Basic Culture Medium DM-160 Amino Acidsmg/l Vi tamins mg/l . .
Ala 400 B~ 1.0 Arg 100 B2 1. 0 A5p 25 B 6 1 . O
Asn 25 B 12 0 . 005 Cys-hCl 80 Pantothenic 1.0 acid Glu 150 Nicotinamide 1.0 Gln 300 Biotin 0.1 Gly 15 Choline HCl 5 . 0 His 30 C 1.0 Ile 150 Folic acid 1.0 Leu 400 Inositol 5.0 Lys 100 Met 80 Phe 80 Pro 12 Ser 8 0 Thr 100 ~rp 40 Tyr 5 0 Val 85 ,~ .
:
Claims (18)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A method for culturing cells or tissue of lower animals comprising culturing a sample of cells or tissue of a lower animal in a nutritive culture medium for cells and tissues of lower animals, said culture medium containing an effective amount of an extract of algae capable of causing cell multiplication of said cells or tissue.
2. A method according to claim 1, in which said algae is selected from the group consisting of Chlorella, Scenedesmus and Spirulina.
3. A method according to claim 2, wherein said extract is produced by extraction of said algae with an aqueous solvent.
4. A method according to claim 2, wherein said extract is produced by extraction of said algae with water at a temperature from 50°C to 150°C, for from 0.5 to 120 minutes.
5. A method according to claim 3, wherein said aqueous solvent contains dissolved therein an additive selected from the group consisting of an acid, a base, and an organic solvent.
6. A method according to claim 2, wherein said extract comprises substances having molecular weights in the range of 1,000 to 1,000,000, which substances include sugars, proteins, polysaccharides and nucleic acids.
7. A method according to claim 3, in which said culture medium contains from 0.3 to 400µg of said extract, calculated on a dry basis, per ml of said culture medium.
8. A method according to claim 7, wherein said extract is produced by extracting said algae with water at a temperature of from 50°C to 150°C, for from 0.5 to 120 minutes, recovering the crude product that was extracted into said water, and then effecting molecular weight fractionation of said crude product to obtain said extract which is free of components derived from said algae having molecular weights of less than 3,000.
9. A method according to claim 7, wherein said animal tissue and animal cells are selected from the group consisting of normal tissue and cancer cells of a lower animal.
10. A method according to claim 7, wherein said algae is Chlorella.
11. A method according to claim 7, wherein said algae is Scenedesmus.
12. A method according to claim 7, wherein said algae is Spirulina.
13. A method as claimed in claim 7, wherein said culture medium further contains animal serum.
14. A method as claimed in claim 13, wherein said culture medium contains up to 1 percent by weight of animal serum.
15. A method for culturing tissues and cells of lower animals, which comprises:
providing a nutrient culture medium containing, per ml of said culture medium, from 0.3 to 400µg, calculated as the solids, of an aqueous extract of an algae selected from the group consisting of Chlorella, Scenedesmus, and Spirulina, said extract having been prepared by extracting said algae with water at a temperature of from 50°C to 150°C, for from 0.5 to 120 minutes, so that said extract contains sugars, proteins, polysaccharides and nucleic acids having molecular weights in the range of from 1,000 to 1,000,000 and exhibits an activity for promoting multiplication of animal cells;
then placing and culturing on said culture medium a sample which is a piece of animal tissue or animal cells obtained from a lower animal, which sample is free of human cells and human tissues.
providing a nutrient culture medium containing, per ml of said culture medium, from 0.3 to 400µg, calculated as the solids, of an aqueous extract of an algae selected from the group consisting of Chlorella, Scenedesmus, and Spirulina, said extract having been prepared by extracting said algae with water at a temperature of from 50°C to 150°C, for from 0.5 to 120 minutes, so that said extract contains sugars, proteins, polysaccharides and nucleic acids having molecular weights in the range of from 1,000 to 1,000,000 and exhibits an activity for promoting multiplication of animal cells;
then placing and culturing on said culture medium a sample which is a piece of animal tissue or animal cells obtained from a lower animal, which sample is free of human cells and human tissues.
16. A method as claimed in claim 15, wherein said mixture is maintained at at least 100°C for at least one minute, and the amount of said extract added to said culture medium is in the range of 1 to 100µg of said extract, calculated as the solids, per ml of said medium.
17. A method according to claim 15, further including the step of fractionating said extract to remove therefrom components having molecular weights of less than 3,000 before adding said extract to said culture medium.
18. A method according to claim 17, further including the step of further fractionating said extract to remove therefrom components having molecular weights in the range of 30,000 to 70,000 before adding said extract to said culture medium.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP55-139,853 | 1980-10-08 | ||
| JP55139853A JPS603827B2 (en) | 1980-10-08 | 1980-10-08 | Cell culture method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1165707A true CA1165707A (en) | 1984-04-17 |
Family
ID=15255053
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000386671A Expired CA1165707A (en) | 1980-10-08 | 1981-09-25 | Method of tissue culture |
Country Status (3)
| Country | Link |
|---|---|
| JP (1) | JPS603827B2 (en) |
| AU (1) | AU546565B2 (en) |
| CA (1) | CA1165707A (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62174024A (en) * | 1986-01-24 | 1987-07-30 | Kurorera Kogyo Kk | Remedy for neurophilic dysfunction |
| JP2520681B2 (en) * | 1986-08-04 | 1996-07-31 | ザ ユニバーシティ オブ ニュー サウス ウェールズ | Biosynthetic human growth hormone products |
| WO2021066113A1 (en) | 2019-10-01 | 2021-04-08 | 学校法人東京女子医科大学 | Production method for composition for cell culturing, composition for cell culturing obtained by same, and cell culturing method using same |
| IL309565A (en) | 2021-06-23 | 2024-02-01 | Univ Tokyo Womens Medical | Method for producing composition for culturing animal cells, composition for culturing animal cells obtained by said method, and method for culturing animal cells using said composition for culturing animal cells |
-
1980
- 1980-10-08 JP JP55139853A patent/JPS603827B2/en not_active Expired
-
1981
- 1981-09-16 AU AU75269/81A patent/AU546565B2/en not_active Ceased
- 1981-09-25 CA CA000386671A patent/CA1165707A/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| AU546565B2 (en) | 1985-09-05 |
| AU7526981A (en) | 1982-04-22 |
| JPS603827B2 (en) | 1985-01-30 |
| JPS5765179A (en) | 1982-04-20 |
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