DK2588083T3 - Biomatrixscaffolds - Google Patents

Biomatrixscaffolds Download PDF

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DK2588083T3
DK2588083T3 DK11801493.5T DK11801493T DK2588083T3 DK 2588083 T3 DK2588083 T3 DK 2588083T3 DK 11801493 T DK11801493 T DK 11801493T DK 2588083 T3 DK2588083 T3 DK 2588083T3
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cells
biomatrix
tissue
medium
scaffold
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DK11801493.5T
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Lola Cynthia Mcadams Ried
Mitsuo Yamauchi
Cai-Bin Cui
Andrew Zhuang Wang
Michael Edward Werner
Yunfang Wang
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Univ North Carolina Chapel Hill
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    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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Claims (13)

1. Fremgangsmåde til fremstilling af et biomatrixscaffold fra pattedyrslevervæv, omfattende: a) at perfundere eller homogenisere levervævet med et første basalmedie, hvor osmolaliteten af det første medie er fra 250 mOsm/kg til 350 mOsm/kg og det første medie er serum-frit og med en neutral pH; derefter b) at perfundere det biologiske væv eller at ekstrahere homogenatet fra trin (a) med en delipiderende buffer omfattende natriumdeoxycholat og phospholipase A2 i det første medie; derefter c) at perfundere vævet eller at ekstrahere homogenatet fra trin (b) med en buffer ved en neutral pH og omfattende en saltkoncentration fra 2,OM NaCI til 5,OM, koncentrationen er valgt for at bevare uopløselige collagener identificeret i det biologiske væv; derefter d) at perfundere vævet eller at ekstrahere homogenatet fra trin (c) med RNase og DNase i en buffer; og derefter e) at vaske vævet eller homogenatet fra trin (d) med et andet medie der har en neutral pH, er serum-frit og har en osmolalitet fra 250 mOsm/kg til 350 mOsm/kg, for derved at fremstille et intakt eller homogeniseret biomatrixscaffold fra det biologiske væv, idet biomatrixscaffoldet bevarer mindst 95% af dets originale collagener og de fleste collagen-associerede matrixkomponenter og matrixbundne vækstfaktorer, hormoner og cytokiner af det biologiske væv.
2. Fremgangsmåde ifølge krav 1, hvor basal-mediet er valgt fra gruppen bestående af RPMI 1640, DME/F12, DME, F12, Waymouths og Williams medie.
3. Fremgangsmåde ifølge et hvilket som helst af krav 1 eller 2, hvor det andet medie omfatter mindst en af bestanddelene der er til stede i interstitiel væske.
4. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor den delipiderende buffer fra trin (b) omfatter fra 20 enheder/L til 50 enheder/L phospholipase A2 og 1% natriumdeoxycholat i det første medie.
5. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor saltkoncentrationen af bufferen fra trin (c) er fra 3,4M NaCI til 3,5M NaCI ved anvendelse til scaffoldfremstilling fra en adult lever og er fra 4,OM NaCI til 4,5M NaCI ved anvendelse til scaffoldfremstilling fra en fosterlever.
6. Fremgangsmåde ifølge et hvilket som helst foregående krav 1, hvor bufferen fra trin (c) endvidere omfatter en proteaseinhibitor, eventuelt hvor proteaseinhibitoren er sojabønnetrypsininhibitor.
7. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor bufferen fra trin (d) endvidere omfatter en proteaseinhibitor, eventuelt hvor proteaseinhibitoren er sojabønnetrypsininhibitor.
8. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor alle medierne og bufferne fra trinnene (a) til (e) er uden en påviselig mængde af et enzym der nedbryder ekstracellulære matrixkomponenter.
9. Fremgangsmåde ifølge et hvilket som helst af de foregående krav endvidere omfattende sterilisering af biomatrixscaffoldet, eventuelt hvor steriliseringstrinnet omfatter gammabestråling af biomatrixscaffoldet.
10. Biomatrixscaffold fremstillet med fremgangsmåde ifølge et hvilket som helst af kravene 1-9.
11. Fremgangsmåde til fremstilling af en cellekultur, omfattende: a) at fremstille et biomatrixscaffold ifølge fremgangsmåden ifølge et hvilket som helst af kravene 1-9; b) at bringe biomatrixscaffoldet fra trin (a) i kontakt med celledyrkningsmediet i et dyrkningsapparat; og c) at pode biomatrixscaffoldet fra trin (b) med celler, for derved at fremstille en cellekultur.
12. Fremgangsmåde ifølge krav 11 hvor trin (b) omfatter at fryse biomatrixscaffoldet fra trin, at fremstille et frosset afsnit af biomatrixscaffoldet som et celledyrkningssubstrat og at bringe celledyrkningssubstratet i kontakt med celledyrkningsmediet i et dyrkningsapparat.
13. Fremgangsmåde ifølge krav 11 hvor trin (b) omfatter at formale biomatricen til et pulver, hvilket eventuelt udføres i en frysemølle ved eller tæt på temperaturer for flydende nitrogen, at coate mindst del af et dyrkningsapparat med pulveret for at fremstille et celledyrkningssubstratum og at bringe celledyrkningssubstratet i kontakt med celledyrkningsmediet i dyrkningsapparatet.
DK11801493.5T 2010-07-02 2011-07-01 Biomatrixscaffolds DK2588083T3 (da)

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US36093910P 2010-07-02 2010-07-02
PCT/US2011/042805 WO2012003450A2 (en) 2010-07-02 2011-07-01 Biomatrix scaffolds

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