CA1269628A - Cell culture microcarrier, method for preparing same and use thereof for cultivating anchorage-dependent cells - Google Patents
Cell culture microcarrier, method for preparing same and use thereof for cultivating anchorage-dependent cellsInfo
- Publication number
- CA1269628A CA1269628A CA000472941A CA472941A CA1269628A CA 1269628 A CA1269628 A CA 1269628A CA 000472941 A CA000472941 A CA 000472941A CA 472941 A CA472941 A CA 472941A CA 1269628 A CA1269628 A CA 1269628A
- Authority
- CA
- Canada
- Prior art keywords
- microcarrier
- cell culture
- positively charged
- basic groups
- dependent cells
- 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 - Lifetime
Links
- 238000004113 cell culture Methods 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims abstract description 18
- 230000001419 dependent effect Effects 0.000 title claims description 11
- 150000004676 glycans Chemical class 0.000 claims abstract description 15
- 239000005017 polysaccharide Substances 0.000 claims abstract description 15
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 13
- 125000004432 carbon atom Chemical group C* 0.000 claims abstract description 9
- 125000001183 hydrocarbyl group Chemical group 0.000 claims abstract 3
- 150000001450 anions Chemical class 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 3
- 150000004804 polysaccharides Polymers 0.000 claims description 2
- 125000004178 (C1-C4) alkyl group Chemical group 0.000 claims 2
- 125000000816 ethylene group Chemical group [H]C([H])([*:1])C([H])([H])[*:2] 0.000 claims 1
- 230000003472 neutralizing effect Effects 0.000 claims 1
- 125000000217 alkyl group Chemical group 0.000 abstract description 5
- 125000003710 aryl alkyl group Chemical group 0.000 abstract description 4
- 125000003118 aryl group Chemical group 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 28
- 229920002307 Dextran Polymers 0.000 description 14
- 239000011324 bead Substances 0.000 description 13
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 6
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- YMDNODNLFSHHCV-UHFFFAOYSA-N 2-chloro-n,n-diethylethanamine Chemical compound CCN(CC)CCCl YMDNODNLFSHHCV-UHFFFAOYSA-N 0.000 description 5
- 229920005654 Sephadex Polymers 0.000 description 5
- 239000012507 Sephadex™ Substances 0.000 description 5
- 239000012091 fetal bovine serum Substances 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 231100000331 toxic Toxicity 0.000 description 5
- 230000002588 toxic effect Effects 0.000 description 5
- 231100000433 cytotoxic Toxicity 0.000 description 4
- 230000001472 cytotoxic effect Effects 0.000 description 4
- 239000007758 minimum essential medium Substances 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 230000003416 augmentation Effects 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- -1 diethylaminoethyl groups Chemical group 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 150000002430 hydrocarbons Chemical group 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 229960005419 nitrogen Drugs 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 2
- 241000867607 Chlorocebus sabaeus Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 229940095679 poly-dex Drugs 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 238000000954 titration curve Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- QLOKJRIVRGCVIM-UHFFFAOYSA-N 1-[(4-methylsulfanylphenyl)methyl]piperazine Chemical compound C1=CC(SC)=CC=C1CN1CCNCC1 QLOKJRIVRGCVIM-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 241000501458 Cultus Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 238000010934 O-alkylation reaction Methods 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002152 alkylating effect Effects 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 150000001449 anionic compounds Chemical class 0.000 description 1
- 230000003190 augmentative effect Effects 0.000 description 1
- 230000000721 bacterilogical effect Effects 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 210000001840 diploid cell Anatomy 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- HKIOYBQGHSTUDB-UHFFFAOYSA-N folpet Chemical group C1=CC=C2C(=O)N(SC(Cl)(Cl)Cl)C(=O)C2=C1 HKIOYBQGHSTUDB-UHFFFAOYSA-N 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 150000002891 organic anions Chemical class 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910001961 silver nitrate Inorganic materials 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 125000000542 sulfonic acid group Chemical group 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 125000001302 tertiary amino group Chemical group 0.000 description 1
- 125000005425 toluyl group Chemical group 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
- C08B37/0021—Dextran, i.e. (alpha-1,4)-D-glucan; Derivatives thereof, e.g. Sephadex, i.e. crosslinked dextran
-
- 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
- C12N5/0068—General culture methods using substrates
- C12N5/0075—General culture methods using substrates using microcarriers
-
- 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
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/70—Polysaccharides
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Cell Biology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- General Engineering & Computer Science (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Cephalosporin Compounds (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19843402927 DE3402927A1 (de) | 1984-01-28 | 1984-01-28 | Zellkulturmikrotraeger, verfahren zu seiner herstellung und seine verwendung zum zuechten von verankerungsabhaengigen zellen |
| DEP3402927.3 | 1984-01-28 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1269628A true CA1269628A (en) | 1990-05-29 |
Family
ID=6226136
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000472941A Expired - Lifetime CA1269628A (en) | 1984-01-28 | 1985-01-25 | Cell culture microcarrier, method for preparing same and use thereof for cultivating anchorage-dependent cells |
Country Status (10)
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6410979A (en) * | 1987-07-03 | 1989-01-13 | Mitsubishi Chem Ind | Microcarrier for cultivating cell |
| MC2098A1 (fr) * | 1988-06-03 | 1991-02-15 | Italfarmaco Spa | Sels de glycosaminoglycanes,procedes pour leur preparation et compositions pharmaceutiques les contenant |
| US5349089A (en) * | 1989-07-07 | 1994-09-20 | National Starch And Chemical Investment Holding Corporation | Reagent for preparing polycationic polysaccharides |
| US5227481A (en) * | 1989-07-07 | 1993-07-13 | National Starch And Chemical Investment Holding Corporation | Cationic polysaccharides and reagents for their preparation |
| DE69228055T2 (de) * | 1991-02-28 | 1999-05-20 | Anticancer Inc., San Diego, Calif. | Ein gewebekulturverfahren für haut im natürlichen zustand |
| DE19954357C2 (de) * | 1999-11-11 | 2002-09-05 | Cosmedica Consulting Gmbh | Zusammensetzung, enthaltend Heteropolysaccharid und perlenförmiges quervernetztes Polysaccharid, sowie deren Verwendung für medizinische/kosmetische Zwecke |
| DE10307925A1 (de) * | 2002-02-25 | 2003-09-04 | Pentax Corp | Träger für eine Zellkultur und Verfahren zur Kultivierung von Zellen |
| US20080020049A1 (en) * | 2005-02-25 | 2008-01-24 | Andrew Darling | Super-sparger microcarrier beads and precision extrusion deposited poly-epsilon-caprolactone structures for biological applications |
| US20090047260A1 (en) * | 2007-08-17 | 2009-02-19 | Wake Forest University Health Sciences | Keratin biomaterials for cell culture and methods of use |
| WO2015042125A2 (en) | 2013-09-19 | 2015-03-26 | The Board Of Trustees Of The Leland Stanford Junior University | Methods and compositions for producing hepatocyte-like cells |
Family Cites Families (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3910819A (en) * | 1974-02-19 | 1975-10-07 | California Inst Of Techn | Treatment of surfaces to stimulate biological cell adhesion and growth |
| JPS6026121B2 (ja) * | 1977-02-28 | 1985-06-21 | 名糖産業株式会社 | 新規デキストラン誘導体及びその製法 |
| FR2400063A1 (fr) * | 1977-08-08 | 1979-03-09 | Pasteur Institut | Procede d'obtention de supports pour cultures cellulaires et supports obtenus |
| US4293654A (en) * | 1977-10-17 | 1981-10-06 | Massachusetts Institute Of Technology | Cell culture microcarriers |
| SE445116B (sv) * | 1979-09-12 | 1986-06-02 | Pharmacia Fine Chemicals Ab | Sett att odla celler pa mikroberare med fibronektinytskikt |
| US4335215A (en) * | 1980-08-27 | 1982-06-15 | Monsanto Company | Method of growing anchorage-dependent cells |
| US4415668A (en) * | 1981-04-20 | 1983-11-15 | President And Fellows Of Harvard College | Cell culture |
| SE8103137L (sv) * | 1981-05-19 | 1982-11-20 | Pharmacia Ab | Polymer med kvartera aminogrupper |
| SE8103138L (sv) * | 1981-05-19 | 1982-11-20 | Pharmacia Fine Chemicals Ab | Mikroberare for odling av forankringsberoende celler |
| IL74259A0 (en) * | 1984-02-06 | 1985-05-31 | Surface Concepts Pty Ltd | Improved method for cell culture |
| SE454518B (sv) * | 1984-05-21 | 1988-05-09 | Statens Bakteriologiska Lab | Forfarande for odling av diploida celler i nervaro av cellulosafibrer |
| US4661407A (en) * | 1985-01-07 | 1987-04-28 | Kms Fusion, Inc. | Glass-surface microcarrier for anchorage-dependent cell cultivation |
| KR950006247B1 (ko) * | 1986-02-28 | 1995-06-12 | 소니 가부시끼가이샤 | 시간축 신장 재생장치 |
| JPS63237782A (ja) * | 1987-03-27 | 1988-10-04 | Teijin Ltd | 付着性動物細胞の培養方法 |
-
1984
- 1984-01-28 DE DE19843402927 patent/DE3402927A1/de active Granted
-
1985
- 1985-01-14 US US06/691,006 patent/US4824946A/en not_active Expired - Fee Related
- 1985-01-22 AT AT85100611T patent/ATE46717T1/de not_active IP Right Cessation
- 1985-01-22 EP EP85100611A patent/EP0150796B1/de not_active Expired
- 1985-01-22 DE DE8585100611T patent/DE3573273D1/de not_active Expired
- 1985-01-25 CA CA000472941A patent/CA1269628A/en not_active Expired - Lifetime
- 1985-01-25 JP JP60013387A patent/JPS60237991A/ja active Pending
- 1985-01-25 PT PT79884A patent/PT79884B/pt unknown
- 1985-01-25 IE IE850175A patent/IE850175L/xx unknown
- 1985-01-25 ES ES539867A patent/ES8602039A1/es not_active Expired
- 1985-01-28 DK DK37985A patent/DK37985A/da not_active Application Discontinuation
-
1989
- 1989-01-04 US US07/293,197 patent/US4910142A/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| DE3402927A1 (de) | 1985-08-08 |
| IE850175L (en) | 1985-07-28 |
| PT79884A (en) | 1985-02-01 |
| ES539867A0 (es) | 1985-11-16 |
| DE3402927C2 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html) | 1989-03-09 |
| ES8602039A1 (es) | 1985-11-16 |
| US4824946A (en) | 1989-04-25 |
| PT79884B (en) | 1986-10-23 |
| DE3573273D1 (en) | 1989-11-02 |
| EP0150796B1 (de) | 1989-09-27 |
| ATE46717T1 (de) | 1989-10-15 |
| US4910142A (en) | 1990-03-20 |
| DK37985D0 (da) | 1985-01-28 |
| EP0150796A3 (en) | 1988-01-07 |
| DK37985A (da) | 1985-07-29 |
| JPS60237991A (ja) | 1985-11-26 |
| EP0150796A2 (de) | 1985-08-07 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKLA | Lapsed |