DK2330889T3 - Forbedret cellesammensætning og fremgangsmåde til fremstilling af samme - Google Patents
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- DK2330889T3 DK2330889T3 DK09789172.5T DK09789172T DK2330889T3 DK 2330889 T3 DK2330889 T3 DK 2330889T3 DK 09789172 T DK09789172 T DK 09789172T DK 2330889 T3 DK2330889 T3 DK 2330889T3
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/10—Preservation of living parts
- A01N1/12—Chemical aspects of preservation
- A01N1/122—Preservation or perfusion media
- A01N1/125—Freeze protecting agents, e.g. cryoprotectants or osmolarity regulators
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
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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
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0605—Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
Landscapes
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- Gynecology & Obstetrics (AREA)
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Claims (26)
1. Fremgangsmåde til fremstilling af en sammensætning omfattende pattedyrsceller, omfattende: (a) at bringe cellerne i kontakt med en opløsning omfattende (1) dextran, maltodextran, trehalose eller hetastivelse; og (2) humant serumalbumin (HSA) for at danne en celle-holdig opløsning; (b) at filtrere den celle-holdige opløsning gennem et 70 mikrometer til 100 mikrometer filter; og (c) hvis den celle-holdige opløsning omfatter mere end 10 ± 3 x 106 celler pr. milliliter, at fortynde cellerne til ikke mere end 10 ± 3 x 106 celler pr. milliliter med en første fortyndingsopløsning omfattende dextran.
2. Fremgangsmåde ifølge krav 1, hvor cellerne kryo præserve res efter trin (c).
3. Fremgangsmåde ifølge krav 1, hvor opløsningen i trin (a) omfatter 10% HSA (vægt/volumen).
4. Fremgangsmåde ifølge krav 1, hvor den første fortyndingsopløsning omfatter 5,5% dextran 40 (vægt/volumen).
5. Fremgangsmåde ifølge krav 1, hvor den første fortyndingsopløsning omfatter 10% HSA (vægt/volumen).
6. Fremgangsmåde ifølge krav 1, hvor opløsningen i trin (a) eller den første fortyndingsopløsning endvidere omfatter et kryobeskyttelsesmiddel.
7. Fremgangsmåde ifølge krav 6, hvor kryobeskyttelsesmidlet i den første fortyndingsopløsning er DMSO.
8. Fremgangsmåde ifølge krav 1 endvidere omfattende efter trin (c): (d) at fortynde den celle-holdige opløsning med en anden fortyndingsopløsning omfattende dextran, maltodextran, trehalose eller hetastivelse men ikke omfattende HSA, for derved at fremstille en sammensætning omfattende pattedyrsceller.
9. Fremgangsmåde ifølge krav 8, hvor den anden fortyndingsopløsning omfatter 5,5% eller 10% dextran 40 (vægt/volumen).
10. Fremgangsmåde ifølge krav 1, hvor filteret er et 70 mikrometer filter eller et 100 mikrometer filter.
11. Fremgangsmåde ifølge kravene 1 til 10, hvor pattedyrscellerne er isolerede humane adhærente placentale celler.
12. Fremgangsmåde ifølge krav 11, omfattende: (a) at opslæmme en flerhed af isolerede humane adhærente placentale celler i en 5,5% dextran 40 (vægt/volumen), 10% humant serumalbumin (HSA) (vægt/volumen) opløsning for at danne en celle-holdig opløsning; (b) at filtrere den celle-holdige opløsning gennem et 70 mikrometer til 100 mikrometer filter; (c) at fortynde den celle-holdige opløsning i 5,5% dextran 40 (vægt/volumen), 10% HSA (vægt/volumen), og 5% DMSO til ikke mere end 10 ± 3 x 106 celler pr. milliliter; (d) at kryopræservere cellerne; (e) at optø cellerne; og (f) eventuelt at fortynde den celle-holdige opløsning 1:1 til 1:11 med 10% dextran 40 (vægt/volumen) for at fremstille sammensætningen.
13. Fremgangsmåde ifølge krav 11, omfattende: (a) at centrifugere en flerhed af isolerede humane adhærente placentale celler for at opsamle cellerne; (b) at genopslæmme cellerne i 5,5% dextran 40 (vægt/volumen); (c) at centrifugere cellerne for at opsamle cellerne; (d) at genopslæmme cellerne i en 5,5% dextran 40 (vægt/volumen) opløsning der omfatter 10% humant serumalbumin (HSA) (vægt/volumen) for at danne en celle-holdig opløsning; (e) at filtrere den celle-holdige opløsning gennem et 70 mikrometer til 100 mikrometer filter; (f) at fortynde den celle-holdige opløsning i 5,5% dextran 40 (vægt/volumen), 10% HSA (vægt/volumen), og 5% DMSO til ikke mere end 10 ± 3 x 106 celler pr. milliliter; (g) at kryopræservere cellerne; (h) at optø cellerne; og (i) eventuelt at fortynde den celle-holdige opløsning 1:1 til 1:11 med 10% dextran 40 (vægt/volumen) for at fremstille sammensætningen.
14. Fremgangsmåde ifølge krav 11, omfattende: (a) at tilvejebringe en flerhed af isolerede humane adhærente placentale celler i en opløsning omfattende 5,5% dextran 40 (vægt/volumen) og 10% humant serumalbumin (HSA) (vægt/volumen) for at danne en opløsning omfattende isolerede humane adhærente placentale celler; (b) at filtrere opløsningen omfattende isolerede humane adhærente placentale celler gennem et 70 mikrometer til 100 mikrometer filter for at fremstille filtrerede isolerede humane adhærente placentale celler; (c) at fortynde de filtrerede isolerede humane adhærente placentale celler med en mængde afen opløsning omfattende 5,5% dextran 40 (vægt/volumen), 10% HSA (vægt/volumen) og 5% dimethylsulfoxid (DMSO) tilstrækkelig til at bringe de filtrerede isolerede humane adhærente placentale celler til 10 ± 3 x 106 celler pr. milliliter; og (d) at fortynde de isolerede humane adhærente placentale celler med 10% dextran 40 (vægt/volumen) i et forhold på ca. 1:1 til ca. 1:11 isolerede humane adhærente placentale cellendextran 40 for at fremstille sammensætningen.
15. Fremgangsmåde ifølge krav 11, omfattende (a) at bringe isolerede humane adhærente placentale celler i kontakt med en opløsning omfattende 5,5% dextran (vægt/volumen) og 10% humant serumalbumin (HSA) (vægt/volumen); (b) at filtrere de isolerede humane adhærente placentale celler gennem et 70 mikrometer til 100 mikrometer filter; og (c) at fortynde de isolerede humane adhærente placentale celler til 10 ± 3 x 106 celler pr. milliliter med en opløsning omfattende 10% dextran 40 (vægt/volumen), hvor sammensætningen ikke omfatter makrocelleklynger, hvor en makrocelleklynge omfatter en aggregering af celler der er større end ca. 150 mikrometer.
16. Fremgangsmåde ifølge et hvilket som helst af kravene 11 til 15, hvor de isolerede humane adhærente placentale celler er CD10+, CD34- og CD105+.
17. Fremgangsmåde ifølge krav 16, hvor CD10+, CD34- og CD105+ cellerne yderligere er CD200+.
18. Fremgangsmåde ifølge krav 17, hvor CD10+, CD34', CD105+ og CD200+ cellerne er enten CD45' eller CD90+.
19. Fremgangsmåde ifølge krav 17, hvor CD10+, CD34', CD105+ og CD200+ cellerne er CD45' og CD90+.
20. Sammensætning opnåelig med fremgangsmåden ifølge et hvilket som helst af kravene 1 til 19.
21. Sammensætning ifølge krav 20, omfattende en flerhed af isolerede humane adhærente placentale celler i en opløsning omfattende 4-10% dextran 40 (vægt/volumen), hvor sammensætningen omfatter mellem 1,0 ±0,3 x 106 celler pr. milliliter til 5,0 ±1,5 x 106 celler pr. milliliter, og hvor sammensætningen ikke omfatter makrocelleklynger, hvor en makrocelleklynge omfatter en aggregering af celler der er større end ca. 150 mikrometer.
22. Sammensætning ifølge krav 21, hvor sammensætningen omfatter færre end ca. 100 mikrocelleklynger pr. 106 celler.
23. Sammensætning ifølge krav 21, hvor de isolerede humane adhærente placentale celler er CD10+, CD34' og CD105+.
24. Sammensætning ifølge krav 23, hvor CD10+, CD34' og CD105+ cellerne yderligere er CD200+.
25. Sammensætning ifølge krav 24, hvor CD10+, CD34', CD105+ og CD200+ cellerne er enten CD45' eller CD90+.
26. Sammensætning ifølge krav 24 hvor CD10+, CD34', CD105+ og CD200+ cellerne er CD45' og CD90+.
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