DK2588027T3 - Biomatrixscaffolds til spredning efter industriel målestok - Google Patents

Biomatrixscaffolds til spredning efter industriel målestok Download PDF

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DK2588027T3
DK2588027T3 DK11801504.9T DK11801504T DK2588027T3 DK 2588027 T3 DK2588027 T3 DK 2588027T3 DK 11801504 T DK11801504 T DK 11801504T DK 2588027 T3 DK2588027 T3 DK 2588027T3
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biomatrix
cells
tissue
medium
scaffold
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DK11801504.9T
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Marsha Lynn Roach
Richard Harold Malavarca
Yunfang Wang
Lola Cynthia Mcadams Reid
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Univ North Carolina Chapel Hill
Marsha Lynn Roach
Richard Harold Malavarca
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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 (12)

1. Fremgangsmåde til fremstilling af et biomatrixscaffold fra pattedyrslevervæv til spredning efter industriel målestok på et dyrkningsapparat, omfattende: a) at perfundere levervævet eller homogenisere levervævet med en buffer omfattende en saltkoncentration fra 3,5M NaCI til 4,5M NaCI, derefter b) at perfundere levervævet eller at ekstrahere homogenatet fra trin (a) med en delipiderende buffer omfattende natriumdeoxycholat og phospholipase A2 i et første medie, hvor osmolaliteten af det første medie er fra 250 mOsm/kg til 350 mOsm/kg og det første medium er serum-frit og med en neutral pH; 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 NaCI, 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-fri og har en osmolalitet fra 250 mOsm/kg til 350 mOsm/kg, for derved at fremstille et intakt eller homogeniseret biomatrixscaffold fra levervævet, 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; f) at fortynde biomatrixscaffoldet i et basal-medie; g) at fryse biomatrixscaffoldet fra (f); h) at pulverisere biomatrixscaffoldet fra (g) ved kryogen formaling til biomatrixpartikler der varierer i størrelse fra 1 pm til 100 pm; i) at optø biomatrixpartiklerne fra (h) i suspension i basal-medie; og j) at dispergere biomatrixpartiklerne fra trin (i) på et dyrkningsapparat, for derved at fremstille et biomatrixscaffold fra biologisk væv til spredning efter industriel målestok på dyrkningsapparat.
2. Fremgangsmåde ifølge krav 1, endvidere omfattende steriliseringstrinnet af biomatrixscaffoldet, eventuelt hvor steriliseringstrinnet udføres med gammabestråling.
3. Fremgangsmåde ifølge et hvilket som helst af krav 1 eller 2, hvor biomatrixscaffoldet fra (f) fortyndes i et forhold på 1:6 i basal-mediet.
4. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor biomatrixpartiklerne fra (i) fortyndes i et forhold på 1:24 i basal-mediet.
5. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor det første medie omfatter salte, mineraler, aminosyrer, vitaminer og sukkere.
6. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor det første medie er et basal-medie, eventuelt hvor basal-mediet er valgt fra gruppen bestående af RPMI 1640, DME/F12, DME, F12, Waymouths og Williams medie.
7. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor det andet medie omfatter mindst en af bestanddelene der er til stede i interstitiel væske.
8. 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.
9. 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.
10. Fremgangsmåde ifølge et hvilket som helst foregående krav 1, hvor bufferen fra trin (c) endvidere omfatter en proteaseinhibitor, eventuelt hvor proteaseinhibitoren er sojabønnetrypsininhibitor.
11. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor bufferen fra trin (d) endvidere omfatter en proteaseinhibitor, eventuelt hvor proteaseinhibitoren er sojabønnetrypsininhibitor.
12. 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.
DK11801504.9T 2010-07-02 2011-07-01 Biomatrixscaffolds til spredning efter industriel målestok DK2588027T3 (da)

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PCT/US2011/042825 WO2012003463A1 (en) 2010-07-02 2011-07-01 Biomatrix scaffolds for industrial scale dispersal

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