DK2681306T3 - Celledyrkningssystem - Google Patents
Celledyrkningssystem Download PDFInfo
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- DK2681306T3 DK2681306T3 DK12751920.5T DK12751920T DK2681306T3 DK 2681306 T3 DK2681306 T3 DK 2681306T3 DK 12751920 T DK12751920 T DK 12751920T DK 2681306 T3 DK2681306 T3 DK 2681306T3
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Claims (17)
1. Celledyrkningssystem (100), der omfatter: en fluidanordning (5) med en fluidkanal (10) forbundet med en fluidkilde (32, 34), hvilken fluidkilde tilfører fluid til fluidkanalen; en membran (20) placeret inde i kanalen mellem membranbæreelementer (22, 24), hvor mindst en del af membranen er fleksibel; en membranspændingsmekanisme (26) koblet til membranbæreelementeme, der er i stand til at bevæge membranbæreelementeme og foranledige membranen til at strække sig langs mindst én dimension af membranen; mindst ét lag af intestinale epitelceller, der omfatter Villi-strukturer bundet til mindst én overflade af membranen; og hvor forskydningsspændingen på fluidet, der strømmer gennem fluidkanalen er mindre end 1,0 dyne/cm2.
2. Fremgangsmåde til dyrkning af intestinale epitelceller, der omfatter: tilvejebringelse af et fluid, der er egnet til at bevare intestinale epitelceller til et celledyrkningssystem, således at fluidet kommer i kontakt med de intestinale epitelceller, hvor celledyrkningssystemet (100) omfatter: en fluidanordning (5) med en fluidkanal (10) forbundet med en fluidkilde (32, 34), hvilken fluidkilde tilfører fluid til fluidkanalen; en membran (20) placeret inde i kanalen mellem membranbæreelementer (22, 24), hvor mindst en del af membranen er fleksibel; en membranspændingsmekanisme (26) koblet til membranbæreelementer, der er i stand til at bevæge membranbæreelementeme og foranledige membranen til at strække sig på en cyklisk måde langs mindst én dimension af membranen; mindst ét lag af intestinale epitelceller bundet til mindst én overflade af membranen; og hvor forskydningsspændingen på fluidet, der strømmer gennem fluidkanalen, er mindre end 1,0 dyne/cm2; og dyrkning af de intestinale epitelceller in vitro mindst indtil Villi-strukturer er evident.
3. System (100) ifølge krav 1 eller fremgangsmåde ifølge krav 2, hvor forskydningsspændingen på fluidet, der strømmer gennem fluidkanalen (10): (i) er fra 0,008 til 0,08 dyne/cm2; (ii) er omtrent 0,018 dyne/cm2; (iii) kan variere over tid; (iv) kan variere over tid fra 0 til 1000 dyne/cm2 og/eller (v) kan variere over tid fra 0,008 til 0,08 dyne/cm2.
4. System (100) ifølge krav 1 eller 3 eller fremgangsmåde ifølge krav 2 eller 3, hvor membranen (20) foranlediges til at strække sig: (i) fra 0 % til 50 %; (ii) fra 5 % til 15 %; (iii) omtrent 10 %; (iv) mere end 15 % for at frembringe en unormal kondition/tilstand for de intestinale epitelceller; (v) på en cyklisk måde ved en hastighed i intervallet fra 0,01 Hz til 2 Hz; (vi) på en cyklisk måde ved en hastighed i intervallet fra 0,05 Hz til 0,25 Hz; (vii) på en cyklisk måde ved en hastighed på 0,15 Hz; (viii) på en cyklisk måde ved en hastighed større end 0,2 Hz for at frembringe en unormal kondition/tilstand for de intestinale epitelceller og/eller (ix) på en uregelmæssig eller intermitterende måde.
5. System (100) ifølge et hvilket som helst af krav 1, 3 eller 4 eller fremgangsmåde ifølge et hvilket som helst af krav 2 til 4, hvor fluidet strømmer gennem fluidkanalen (10) ved en strømningshastighed på: (i) mindre end 500 pL/h; (ii) mindre end 100 pL/h; (iii) fra 0 til 50 pL/h og/eller (iv) omtrent 30 pL/h.
6. System (100) ifølge et hvilket som helst af krav 1 eller 3 til 5 eller fremgangsmåde ifølge et hvilket som helst af krav 2 til 5, der endvidere omfatter mindst én type afbindingsmolekyle, der understøtter adhæsion af en flerhed af levende celler, der coater mindst én side af membranen (20).
7. System (100) ifølge krav 6 eller fremgangsmåde ifølge krav 6, hvor det mindst ene bindingsmolekyle er valgt fra gruppen bestående af: collagen; collagen type I; MATRIGEL™; ekstracellulær matrix; laminin; proteoglycan; vitronectin; fibronection; poly-D-lysin; polypeptider; oligonukleotider; DNA og polysaccharid.
8. System (100) ifølge et hvilket som helst af krav 1 eller 3 til 7 eller fremgangsmåde ifølge et hvilket som helst af krav 2 til 7, hvor de intestinale epitelceller er valgt fra gruppen bestående af: Caco2-celler; HT-29-celler; primære tyndtarmsepitelceller; primære tyktarmsepitelceller; iPS-celler; ESC-celler; stamceller; panetheceller; krypt-celler og mucus-sekreterende celler.
9. System (100) ifølge et hvilket som helst af krav 1 eller 3 til 8 eller fremgangsmåde ifølge et hvilket som helst af krav 2 til 8, hvor systemet endvidere omfatter mindst ét lag af endotelceller på mindst den anden overflade af membranen (20).
10. System (100) ifølge et hvilket som helst af krav 1 eller 3 til 9 eller fremgangsmåde ifølge et hvilket som helst af krav 2 til 9, hvor systemet endvidere omfatter mikrobielle celler eller patogener.
11. System (100) ifølge krav 10 eller fremgangsmåde ifølge krav 10, hvor de mikrobielle celler er valgt fra gruppen bestående af: lactobacillus; bacterioider; ruminococcus; Ppptococcus; peptostreptococcus; bifidobacterium; escherichia; achromobacter; Acidaminococcus fermentans; Acinetobacter cacoaceticus; Aeromonas; Alcaligenes faecalis; Bacillus; Butyriviberio fibrosolvens; Camplyobacter; Campylobacter coli; Clostridium difficile; Clostridium sordelli; Enterobacter cloacae; Enterococcus faecalis; Enterococcus faecium; Escherichia coli; Flavobacterium; Mycobacterium; Mycoplasma; Plesiomonas shigelloides; Propionibacterium aenes; Pseudomonas aeruginosa; Ruminococcus bromii; Sarcina; Staphylococcus aureus; Streptococcus anginosus; Veillonella; Vibrio; Yersinia enterocolitica; Lactobacillus rhamnosus; Lactobacillus rhamnosus GG; Bifidobacterium breve; Bifidobacterium longum; Bifidobacterium infantis; Lactobacillus acidophilus; Lactobacillus plantarum; Lactobacillus paracasei; Lactobacillus bulgaricus og Streptococcus thermophilus.
12. System (100) ifølge krav 10 eller 11 eller fremgangsmåde ifølge krav 10 eller 11, hvor patogeneme er valgt fra gruppen bestående af: enterotoksigene Escherichia coli; Bilophila wadsworthia', Shigella; Yersinia; Pleisiomonas; Vibrio; Aeromonas; Campylobacter; Crytosporidia; Coccidosis; Salmonella; Helicohacter pylori; Clostridium difficile; Salmonella kedougow, bacteroider; clostridium; firmicutes; Shigellia dysenteriae; Salmonella enterica; Salmonella typhi; Listeria; Listeria monocytogenes; Vibrio parahaemolyticus; Proteus; Vibrio cholerae; Enterococcus faecalis; Yersinia enterocolitica og Campylobacter jejuni; rotavirus; norwalk-lignende vira; adenovira; astrovira; sapporo-lignende vira; torovira; coronavira; picomavira; herpes vira; norovira; Candida; Aspergillus; Candida albicans; enkeltcellede parasitter; flercellede parasitter; amøber; orm; bændelorm; protozoer; ikter; rundorme; bømeorm; hageorm; Giradla lamblia; Cryptosporidium og Entamoeba histolytica.
13. System (100) ifølge et hvilket som helst af krav 10 til 12 eller fremgangsmåde ifølge et hvilket som helst af krav 10 til 12, hvor de mikrobielle celler (i) er aerobe; (ii) er anaerobe eller (iii) omfatter både aerobe og anaerobe mikrobielle celler; og/eller (iv) er til stede i den første celledyrkningskanal.
14. System (100) ifølge et hvilket som helst af krav 1 eller 3 til 13 eller fremgangsmåde ifølge et hvilket som helst af krav 2 til 13, hvor membranen (20) er mindst delvist porøs.
15. System (100) ifølge et hvilket som helst af krav 1 eller 3 til 14 eller fremgangsmåde ifølge et hvilket som helst af krav 2 til 14, hvor systemet er forbundet eller sammenkoblet med et andet celledyrkningssystem, der omfatter celler eller væv, der ikke er af intestinal oprindelse; fortrinsvis hvor det andet celledyrkningssystem omfatter leverceller eller -væv.
16. System (100) ifølge krav 1, hvor membranen (20) adskiller kanalen (10) i en første celledyrkningskanal (12) og en anden celledyrkningskanal (14).
17. System (100) ifølge krav 16, hvor fluidet tilført fluidkanalen (10) foranlediges til at strømme gennem én eller flere af den første celledyrkningskanal (12) og den anden celledyrkningskanal (14).
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Application Number | Priority Date | Filing Date | Title |
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US201161447540P | 2011-02-28 | 2011-02-28 | |
PCT/US2012/026934 WO2012118799A2 (en) | 2011-02-28 | 2012-02-28 | Cell culture system |
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DK2681306T3 true DK2681306T3 (da) | 2019-04-23 |
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