DK1517740T3 - Tangentialflowfiltreringsenheder og fremgangsmåder til leukocytberigelse - Google Patents
Tangentialflowfiltreringsenheder og fremgangsmåder til leukocytberigelse Download PDFInfo
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- DK1517740T3 DK1517740T3 DK03761157.1T DK03761157T DK1517740T3 DK 1517740 T3 DK1517740 T3 DK 1517740T3 DK 03761157 T DK03761157 T DK 03761157T DK 1517740 T3 DK1517740 T3 DK 1517740T3
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- leukocytes
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- dendritic cells
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- A61M1/3496—Plasmapheresis; Leucopheresis; Lymphopheresis
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- B—PERFORMING OPERATIONS; TRANSPORTING
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Claims (58)
1. Tangentialflowfiltreringsenhed til fremstilling af en cellepopulation, som er beriget med leukocytter, ved selektiv fjernelse af blodkomponenter såsom plasma, trombocytter og erytrocytter, hvilken filtreringsenhed omfatter: en udskillerenhed (1) med et cylindrisk tværstrømskammer (3) og et filtratkammer (4), der er adskilt af et cirkulært filter (5), som er anbragt derimellem, hvilket filter (5) er i væskeforbindelse med tværstrømskammeret (3) og filtratkammeret (4); hvor filterdiameteren i alt væsentligt er den samme som diameteren af tværstrømskammeret; hvor det cylindriske tværstrømskammer (3) har et indløb (6) og et udløb (7), hvor indløbet (6) er anbragt i umiddelbar nærhed af filterets retentatoverflade med henblik på at føre en prøve af blodkomponenter, som omfatter leukocytter, ind i tværstrømskammeret (3) og parallelt med filterets (5) retentatoverflade; og udløbet (7) er beliggende i umiddelbar nærhed af filterets (5) midte og vinkelret på filterets (5) retentatoverflade, en pumpe (14) til at pumpe væske ind i tværstrømskammeret (3) gennem indløbet (6), hvilken pumpe er udformet til at indføre væsken i det cylindriske tværstrømskammer med en tilførselshastighed på 5 til 100 gange filtreringshastigheden; hvor dette arrangement får væsken til at strømme i en indadgående spiral i retning mod filterets midte, så der frembringes en hvirvelbevægelse i væsken, og hvor filteret (5) har en gennemsnitlig porestørrelse i intervallet fra 3 til 8 mikron, således at strømning af prøven gennem filteret (5) beriger prøven af blodkomponenter med leukocytter.
2. Enhed ifølge krav 1, som endvidere omfatter: en genindvindingsenhed (2), der omfatter et indløb (10) og et udløb (11), hvor tværstrømskammeret (3) og genindvindingsenheden (2) er indbyrdes forbundet i et sløjfearrangement, hvor tværstrømskammerets indløb (6) er i væskeforbindelse med genindvindingsenhedens udløb (11), og tværstrømskammerets udløb (7) er i væskeforbindelse med genindvindingsenhedens indløb (10).
3. Enhed ifølge krav 2, hvor genindvindingsenheden endvidere omfatter et prøveindløb (12) og et vaskeindløb (13).
4. Enhed ifølge krav 3, som endvidere omfatter en kilde til erstatningsvæske, som er i væskeforbindelse med vaskeindløbet (13).
5. Enhed ifølge krav 4, hvor erstatningsvæsken er en isotonisk buffer eller vævsdyrkningsmedium.
6. Enhed ifølge et hvilket som helst af kravene 1 -5, som endvidere omfatter et cellebehandlingsapparat, der er i væskeforbindelse med udskillerenheden.
7. Enhed ifølge krav 6, hvor cellebehandlingsapparatet omfatter perler.
8. Enhed ifølge krav 6 eller 7, hvor cellebehandlingsapparatet omfatter et middel til dyrkning af cellepopulationen, som er beriget med leukocytter.
9. Enhed ifølge krav 8, hvor midlet til dyrkning omfatter: en beholder med en første port og en anden port; et substrat til binding af monocytiske forstadier til dendritiske celler, hvilket substrat er i væskeforbindelse med den første port og den anden port; en sigte til at tilbageholde substratet i beholderen, hvilken sigte har en porestørrelse, som er tilstrækkelig til at tillade passage af monocytiske forstadier til dendritiske celler og dendritiske celler derigennem; en aftapningsledning, som er i væskeforbindelse med den første port; og en opsamlingsledning, som er i væskeforbindelse med den første port.
10. Enhed ifølge krav 9, som endvidere omfatter en flerhed af væskekilder, der er i væskeforbindelse med den første port eller den anden port.
11. Enhed ifølge krav 9 eller 10, som endvidere omfatter en forseglelig vævsdyrkningsbeholder, der er tilpasset til aseptisk modtagelse af de monocytiske forstadier til dendritiske celler.
12. Enhed ifølge krav 11, hvor den forseglelige vævsdyrkningsbeholder er en vævsdyrkningspose, -kolbe eller -bioreaktor.
13. Enhed ifølge et hvilket som helst af kravene 10-12, hvor væskekilderne omfatter bindemedier, vaskebuffer og elueringsbuffer.
14. Enhed ifølge et hvilket som helst af kravene 10-13, som endvidere omfatter en pumpe, der er i væskeforbindelse med flerheden af væskekilder og den første port.
15. Enhed ifølge et hvilket som helst af kravene 1 -14, som endvidere omfatter: et temperaturreguleringsmiddel til at bibeholde substratet ved en forudbestemt temperatur.
16. Enhed ifølge krav 15, hvor temperaturreguleringsmidlet er et varmeapparat.
17. Enhed ifølge et hvilket som helst af kravene 1 -16, hvor filterets porestørrelse er fra ca. 3 mikron til ca. 7 mikron eller fra ca. 3 mikron til ca. 5,5 mikron.
18. Enhed ifølge et hvilket som helst af kravene 1 -17, som endvidere omfatter: en kilde til blodkomponenter, der er i væskeforbindelse med tværstrømskammerets indløb.
19. Enhed ifølge krav 18, hvor kilden til blodkomponenter er en leukofereseenhed.
20. Tangentialflowsenhed ifølge krav 1 til berigelse af en prøve af blodkomponenter med leukocytter, hvor enheden omfatter et middel til reduktion af en filtreringshastighed (15) gennem filteret; og hvor filteret (5) har en porestørrelse på fra 3 mikron til 7 mikron.
21. Enhed ifølge krav 20, som endvidere omfatter et middel til tilvejebringelse af en forudbestemt koncentration af blodceller i prøven, hvor den forudbestemte koncentration af blodceller er 107 til 1010 celler pr. milliliter.
22. Enhed ifølge et hvilket som helst af kravene 1 -19, hvor tværstrømskammeret forefindes over filteret og filtratkammeret.
23. Enhed ifølge et hvilket som helst af kravene 1 -19, hvor tværstrømskammeret forefindes under filteret og filtratkammeret.
24. Fremgangsmåde til separation af leukocytter fra en prøve af blodkomponenter fra et individ, hvor prøven omfatter leukocytter, hvilken fremgangsmåde omfatter: (i) at indføre prøven i en udskillerenhed (1) i enheden ifølge et hvilket som helst af kravene 1 -21 gennem et indløb (6) i udskillerenheden; (ii) at underkaste prøven tværstrømning, som i alt væsentligt er parallel med et filter (5) med en porestørrelse på fra ca. 3 til ca. 8 mikron; (iii) at underkaste væsken filtrering gennem filteret (5); og (iv)at fjerne ikke-leukocyt-blodkomponenter selektivt fra prøven for at danne en cellepopulation, der er beriget med leukocytter.
25. Fremgangsmåde ifølge krav 24, som endvidere omfatter: at klargøre prøven fra individet ved leukoferese, densitetscentrifugering, differentiallyse, filtrering eller frembringelse af et leukocyt-trombocyt-lag til indførelse i udskillerenheden (1).
26. Fremgangsmåde ifølge krav 24 eller 25, hvor ikke-leukocyt-blodkomponenterne indbefatter plasma, trombocytter og/eller erytrocytter.
27. Fremgangsmåde ifølge et hvilket som helst af kravene 24-26, som endvidere omfatter at gentage trin (i), (ii) og (iii) mindst to gange for at danne en cellepopulation, der er beriget med leukocytter.
28. Fremgangsmåde ifølge et hvilket som helst af kravene 24-26, hvor den berigede cellepopulation omfatter mindst 20% leukocytter eller mindst 60% leukocytter.
29. Fremgangsmåde ifølge et hvilket som helst af kravene 24-28, som endvidere omfatter at vaske cellepopulationen, der er beriget med leukocytter, med en vaskeopløsning.
30. Fremgangsmåde ifølge et hvilket som helst af kravene 24-29, som endvidere omfatter at fremstille monocytiske forstadier til dendritiske celler ud fra cellepopulationen, der er beriget med leukocytter.
31. Fremgangsmåde ifølge krav 30, hvor isoleringen af monocytiske forstadier til dendritiske celler omfatter: at etablere kontakt mellem et substrat til binding af monocytiske forstadier til dendritiske celler og cellepopulationen, der er beriget med leukocytter; at lade monocytiske forstadier til dendritiske celler i cellepopulationen binde sig reversibelt til substratet for at danne komplekser, der omfatter monocytiske forstadier til dendritiske celler og substrat; at separere komplekserne fra de ikke-bindende leukocytter for at opnå komplekser, der omfatter monocytiske forstadier til dendritiske celler; og at dyrke de monocytiske forstadier til dendritiske celler for at differentiere forstadierne for at danne umodne eller modne dendritiske celler.
32. Fremgangsmåde ifølge krav 31, hvor de monocytiske forstadier til dendritiske celler elueres fra substratet inden dyrkning eller dyrkes på substratet.
33. Fremgangsmåde ifølge krav 32, hvor substratet omfatter mikroperler af glas, polystyren, plast eller glasbelagt polystyren.
34. Fremgangsmåde til berigelse af en prøve af blodkomponenter med leukocytter, hvilken fremgangsmåde omfatter: 1) at indføre prøven i en tangentialflowfiltreringsenhed (TFF-enhed) ifølge et hvilket som helst af kravene 1 -21; 2) at recirkulere prøven gennem TFF-enheden med en forudbestemt tilførselshastighed og en forudbestemt filtreringshastighed, hvor den forudbestemte tilførselshastighed er mindst fem gange større end den forudbestemte filtreringshastighed: hvor den forudbestemte filtreringshastighed er mindre end den uhindrede filtreringshastighed for filteret; og 3) at isolere en cellepopulation, der er beriget med leukocytter.
35. Fremgangsmåde ifølge krav 34, hvor den berigede cellepopulation er i alt væsentligt fri for ikke-leukocyt-blodkomponenter.
36. Fremgangsmåde ifølge krav 34, som endvidere omfatter: at indsamle blod fra et individ og klargøre prøven fra blodet ved leukoferese, densitetscentrifugering, differentiallyse, filtrering eller frembringelse af et leukocyt-trombocyt-lag.
37. Fremgangsmåde ifølge et hvilket som helst af kravene 34-36, hvor ikke-leukocyt-blodkomponenterne indbefatter plasma, trombocytter og/eller erytrocytter.
38. Fremgangsmåde ifølge et hvilket som helst af kravene 34-37, hvor den berigede cellepopulation omfatter mindst ca. 20% leukocytter eller mindst ca. 60% leukocytter.
39. Fremgangsmåde ifølge et hvilket som helst af kravene 34-38, som endvidere omfatter at vaske den berigede cellepopulation med en vaskeopløsning.
40. Fremgangsmåde ifølge et hvilket som helst af kravene 34-39, som endvidere omfatter at fremstille dendritiske celler ud fra den berigede cellepopulation.
41. Fremgangsmåde ifølge krav 40, hvor de dendritiske celler fremstilles ved: at etablere kontakt mellem et substrat til binding af monocytiske forstadier til dendritiske celler og den berigede cellepopulation; at lade monocytiske forstadier til dendritiske celler i den berigede cellepopulation binde sig reversibelt til substratet for at danne komplekser, der omfatter monocytiske forstadier til dendritiske celler og substrat; at separere komplekserne fra de ikke-bindende leukocytter for at opnå komplekser, der omfatter monocytiske forstadier til dendritiske celler; og at dyrke de monocytiske forstadier til dendritiske celler for at differentiere forstadierne for at danne umodne eller modne dendritiske celler.
42. Fremgangsmåde ifølge krav 41, hvor substratet omfatter mikroperler af glas, polystyren, plast eller glasbelagt polystyren.
43. Fremgangsmåde ifølge krav 41 eller 42, som endvidere omfatter at isolere de umodne eller modne dendritiske celler.
44. Fremgangsmåde ifølge krav 37, hvor monocytterne dyrkes med cytokiner, som fremmer differentieringen af monocytter til dendritiske celler.
45. Fremgangsmåde ifølge krav 44, hvor cytokinerne er GM-CSF, GM-C-SF og IL-4.
46. Fremgangsmåde ifølge krav 44 eller 45, hvor de dendritiske celler modnes til modne dendritiske celler.
47. Fremgangsmåde ifølge krav 45, hvor de dendritiske celler dyrkes med et antigen under betingelser, som er befordrende for omsætning af antigenet til dannelse af antigenbærende dendritiske celler.
48. Fremgangsmåde ifølge et hvilket som helst af kravene 34-47, hvor filteret har en porestørrelse på fra 3 til 5,5 mikron.
49. Fremgangsmåde ifølge krav 48, hvor leukocytterne omfatter CD34+-celler.
50. Fremgangsmåde ifølge krav 48 eller 49, hvor prøven af blodkomponenter er fra en donor, som er blevet behandlet med mindst ét stamcellemobiliserende middel.
51. Fremgangsmåde ifølge krav 50, hvor det stamcellemobiliserende middel er G-CSF eller cyclophosphamid.
52. Fremgangsmåde ifølge et hvilket som helst af kravene 48-51, som endvidere omfatter at berige leukocytterne med CD34+-cellerne.
53. Fremgangsmåde ifølge krav 52, hvor berigelsen af leukocytter med CD34+-cellerne omfatter at anvende et anti-CD34-antistof, som er konjugeret til magnetiske perler.
54. Fremgangsmåde ifølge et hvilket som helst af kravene 48-53, som endvidere omfatter at ekspandere CD34+-cellerne ex vivo.
55. Fremgangsmåde ifølge et hvilket som helst af kravene 48-51, som endvidere omfatter at fremstille monocytafledte pluripotente stamceller ud fra cellepopulationen, der er beriget med leukocytter.
56. Fremgangsmåde ifølge krav 55, som endvidere omfatter at inducere differentiering af en progenitor- eller stamcelle.
57. Fremgangsmåde ifølge krav 48, som endvidere omfatter at inducere transdifferentiering af en differentieret celle.
58. Anvendelse af en tangentialflowfiltreringsenhed ifølge et hvilket som helst af kravene 1-21 til fremstilling af en cellepopulation, som er beriget med leukocytter, ved selektiv fjernelse af blodkomponenter såsom plasma, trombocytter og erytrocytter.
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| US39073002P | 2002-06-19 | 2002-06-19 | |
| PCT/US2003/019428 WO2004000444A1 (en) | 2002-06-19 | 2003-06-19 | Tangential flow filtration devices and methods for leukocyte enrichment |
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| DK03761157.1T DK1517740T3 (da) | 2002-06-19 | 2003-06-19 | Tangentialflowfiltreringsenheder og fremgangsmåder til leukocytberigelse |
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| WO2003080801A2 (en) * | 2002-03-19 | 2003-10-02 | Advanced Research & Technology Transfer | Adipose stromal stem cells for tissue and vascular modification |
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- 2003-06-19 RU RU2005101318/14A patent/RU2328317C2/ru active IP Right Revival
- 2003-06-19 US US10/517,871 patent/US7695627B2/en active Active
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