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IMMUNE INFORMED-MUCIN HYDROGELS AS LOCAL IMMUNE ISOLATION FOR ENCAPSULATED ISLET TRANSPLANTATION: IN VITRO FEASIBILITY VALIDATION
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH).
2019 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Within the type 1 diabetic research several applications have been evaluated to improve lifequality among type 1 diabetic patients. The aim of this project was to validate in vitro thefeasibility of using mucin hydrogel for islet transplantation . The project started with macroencapsulation of Min6-M9 cancer cell line extracted from 13 weeks old mice. The projectcontinued with encapsulation of Min6-M9 cells in microscopic hydrogel using a microfluidicdevice to evaluate the possibility to automate the encapsulation process, but also to facilitatethe future transplantation process upon surgery due to practical reasons since microscopicmucin hydrogel has smaller diameter than macro-encapsulated mucin hydrogel. Finally,primary islets isolated from 4 months old C57B1/6J laboratory mice was obtained and macroencapsulated in mucin hydrogel. In the experiments the samples were cultivated long termand continuously tested through quantitative data assay e.g. insulin secretion assay andAlamar Blue assay followed with qualitative data analysis through e.g. bright field imagesand measurements of the spherical diameters from single cells to clusters. This was followedwith LIVE/DEAD staining. From the obtained data from both Min6-M9 inmacro/microscopic hydrogel and primary islets it was possibility to show the long term isletssurvival of encapsulated Min6-M9 cells and primary islets. In addition, all samples secretedinsulin wherein primary islets had a clear insulin response upon glucose stimulation. Fromthe experiments in vitro the conclusion can be made that mucin hydrogel do supportencapsulated islets long term function and survival. From this, the feasibility usingencapsulated islets in mucin hydrogel needs to be investigated in vivo.

Abstract [sv]

Inom Typ-1 Diabetes forskningen har flera applikationer blivit utvärderade för att förbättralivskvaliteten hos typ-1 Diabetes patienter. Detta projekt är en av dessa medicinskaapplikationer som inneburit att utvärdera möjligheten att använda mucin hydrogel för islettransplantation in vitro . Projektet startade med en makro-inkapsling av Min6-M9 cancercellerfrån 13 veckor gamla möss. Projektet fortsatte med inkapsling av Min6-M9 i en mikroskopiskmucin hydrogel genom att använda sig av en mikrofluidik utrustning för att utvärderamöjligheten till en automationsprocess av inkapslingen, men också av praktiska skälunderlätta framtida transplantations processer under operation då en mikroskopisk mucinhydrogel har lägre diameter än en makroskopisk mucin hydrogel. Slutligen, användesprimary islets från 4 månader gamla C57B1/6J laboratoriemöss varpå dessa inkapslades i enmucin hydrogel. I alla experiment kultiverades proverna under en längre tid varpå demtestades genom kvantitativa dataanalyser e.g. insulin sekretion assay och Alamar Blue assayföljt med kvalitativa dataanalyser genom mikroskopisk fotografering och mätningar avdiameterna av enstaka celler till kluster. Detta åtföljdes av LIVE/DEAD staining. Frånerhållna data av inkapslade Min6-M9 i både makroskopisk/mikroskopisk hydrogel ochprimary islets var det möjligt att dra slutsatsen att proverna kunde överleva under längre tid.Utöver detta, påvisade alla prover en insulinsekretion varpå primary islet hade en tydliginsulinrespons i anslutning till glukos stimulering. Från experimenten utförda in vitro kanslutsatsen dras att mucin hydrogel ger support till inkapslade islets funktion och överlevnadoch därmed bör möjligheten in vivo närmre utvärderas.

Place, publisher, year, edition, pages
2019.
Series
TRITA-CBH-GRU ; 2020:030
Keywords [en]
Type-1 Diabetes; Hydrogel; Encapsulation; Mucin; Min6-M9; Primary islets; Alginate; FRET; Alamar Blue assay
Keywords [sv]
Typ-1 Diabetes; Hydrogel; Inkapsling; Mucin; Min6-M9; Primary islets; Alginate; FRET; Alamar Blue assay
National Category
Medical Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-271655OAI: oai:DiVA.org:kth-271655DiVA, id: diva2:1421348
Educational program
Master of Science in Engineering - Biotechnology
Examiners
Available from: 2020-04-02 Created: 2020-04-02 Last updated: 2020-04-02Bibliographically approved

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