DK2601288T3 - Forenklede basemedier til human pluripotent cellekultur - Google Patents

Forenklede basemedier til human pluripotent cellekultur Download PDF

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DK2601288T3
DK2601288T3 DK11751704.5T DK11751704T DK2601288T3 DK 2601288 T3 DK2601288 T3 DK 2601288T3 DK 11751704 T DK11751704 T DK 11751704T DK 2601288 T3 DK2601288 T3 DK 2601288T3
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cells
cell
human
pluripotent stem
medium
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DK11751704.5T
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James A Thomson
Guokai Chen
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Wisconsin Alumni Res Found
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Claims (16)

1. Defineret albuminfrit medium omfattende følgende bestanddele: vand, salte, aminosyrer, vitaminer, en carbonkilde, insulin, FGF2, selen, transferrin og en blandt TGF-β og NODAL, hver kombineret i en mængde, der understøtter proliferation af humane pluripotente stamceller.
2. Fremgangsmåde til dyrkning af humane pluripotente stamceller, hvilken fremgangsmåde omfatter trinnene: at anbringe humane pluripotente stamceller på en matrix; og at bringe de humane pluripotente stamceller i kontakt med det definerede albuminfri medium ifølge krav 1.
3. Fremgangsmåde ifølge krav 2, hvor matricen omfatter laminin eller vitronectin.
4. Fremgangsmåde ifølge krav 2 eller 3, hvor de humane pluripotente stamceller bringes i kontakt med mediet under hypoksiske betingelser.
5. Fremgangsmåde ifølge et hvilket som helst af kravene 2 til 4, hvor de humane pluripotente stamceller er humane embryoniske stamceller eller humane inducerede pluripotente stamceller.
6. Fremgangsmåde til kloning af en human pluripotent stamcelle, hvilken fremgangsmåde omfatter trinnene: at anbringe humane pluripotente stamceller ved kloningsdensitet i et defineret albuminfrit medium omfattende følgende bestanddele: vand, salte, aminosyrer, vitaminer, en carbonkilde, insulin, FGF2, selen, transferrin og en blandt TGF-β og NODAL, hver kombineret i en mængde, der understøtter kloning af humane pluripotente stamceller.
7. Fremgangsmåde ifølge krav 6, hvor mediet yderligere omfatter en ROCK-inhibitor og/eller blebbistatin.
8. Fremgangsmåde ifølge krav 7, hvor ROCK-inhibitoren er udvalgt fra gruppen bestående af HA100 og Y27632.
9. Fremgangsmåde til afledning af en human induceret pluripotent stamcelle (iPS-celle) under definerede betingelser, hvilken fremgangsmåde omfatter trinnet: at reprogrammere en human somatisk celle i et defineret albuminfrit medium omfattende følgende bestanddele: vand, salte, aminosyrer, vitaminer, en carbonkilde, insulin, FGF2, selen, transferrin og en blandt TGF-β og NODAL, enkelt kombineret i en mængde, der understøtter reprogrammering af humane somatiske celler til udledning af en human IPS-celle.
10. Fremgangsmåde ifølge krav 9, hvor reprogrammeringstrinnet omfatter: (a) at bringe den humane somatiske celle i kontakt med TGF-β i 5-10 dage; (b) at bringe den humane somatiske celle i kontakt med hydrocortison; eller (c) at bringe den humane somatiske celle i kontakt med butyrat.
11. Fremgangsmåde ifølge krav 10, yderligere omfattende trinnene: at fjerne TGF-β efter 5-10 dage; og at bringe den humane somatiske celle i kontakt med et medium i det væsentlige bestående af: vand, salte, aminosyrer, vitaminer, en carbonkilde, insulin, FGF2, selen og transferrin.
12. Fremgangsmåde ifølge et hvilket som helst af kravene 9 til 11, hvor iPS-cellen er afledt under xenofri betingelser.
13. Fremgangsmåde ifølge et hvilket som helst af kravene 9 til 12, hvor re-programmingstrinnet omfatter at anvende en virusvektor eller omfatter at anvende en episomvektor.
14. Defineret albuminfrit vækstmedium, der understøtter langtidsdyrkning af humane pluripotente stamceller, hvilket medium i det væsentlige består af følgende bestanddele: vand, salte, aminosyrer, vitaminer, en carbonkilde, insulin, FGF2, selen, transferrin og en blandt TGF-β og NODAL, hver bestanddel kombineret i en mængde, der er tilstrækkelig til at understøtte langtidsdyrkning af humane pluripotente stamceller.
15. In wYro-cellekultursammensætning af humane pluripotente stamceller i det definerede albuminfri medium ifølge krav 1 eller 14.
16. Fremgangsmåde til kryopræservering af humane pluripotente stamceller, hvilken fremgangsmåde omfatter trinnet: at fryse humane pluripotente stamceller i det definerede albuminfri medium ifølge krav 1 eller 14.
DK11751704.5T 2010-08-05 2011-08-05 Forenklede basemedier til human pluripotent cellekultur DK2601288T3 (da)

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US37112810P 2010-08-05 2010-08-05
PCT/US2011/046796 WO2012019122A2 (en) 2010-08-05 2011-08-05 Simplified basic media for human pluripotent cell culture

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US (1) US9279103B2 (da)
EP (1) EP2601288B1 (da)
JP (2) JP6043999B2 (da)
KR (1) KR101829488B1 (da)
CN (2) CN103180434A (da)
AU (1) AU2011285531B2 (da)
BR (2) BR112013002811A8 (da)
CA (1) CA2807418C (da)
DK (1) DK2601288T3 (da)
IL (1) IL224431A (da)
SG (1) SG187699A1 (da)
WO (1) WO2012019122A2 (da)

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