Skeletspierweefsel engineering: in vivo evaluatie van een prevascularisatie strategie en verkennen van nieuwe synthetische draagstructuren

Lisanne Terrie
In ons labo worden skeletspieren aangemaakt gebaseerd op een fibrine hydrogel. Deze artificiële spieren kunnen mogelijks tegemoetkomen aan een onbeantwoorde medische vraag, namelijk volumetrisch spierverlies, maar kennen meerdere potentiële toepassingen. Ondanks de vooruitgang geboekt in dit veld, zijn er blijvende uitdagingen waarvoor in deze thesis mogelijke oplossingen verder werden bestudeerd.

Kweekspieren uit het lab: toekomstmuziek of realiteit?

Organen en weefsels bouwen in een labo: het klinkt futuristisch, maar toch krijgt dit meer en meer vorm. Dit zou natuurlijk een enorme stap voorwaarts zijn in de geneeskunde, want orgaandonoren zijn beperkt. Met behulp van nieuwe weefselbouw technologie wordt het misschien mogelijk om van onze eigen stamcellen organen te maken in een laboratorium. Hiermee zouden de huidige transplantatie problemen zoals het afstotingsgevaar kunnen omzeild worden. Wij doen onderzoek naar het maken van spieren.

Schade bestrijden met schade?

Ons lichaam bestaat voor ongeveer 40 % uit skeletspieren, die nodig zijn voor beweging. Schade aan deze spieren heeft dan ook een enorme invloed op onze levenskwaliteit. Gelukkig kan een skeletspier zichzelf herstellen na kleine letsels. Dit is mogelijk door de aanwezigheid van spierstamcellen die worden geactiveerd bij schade. Ze kunnen uitgroeien tot nieuwe spiervezels en zo de verloren spiermassa herstellen. Dit systeem schiet echter tekort in geval van groot spierverlies, bijvoorbeeld bij een verkeersongeval. De huidige aanpak van groot spierverlies is het transplanteren van een stuk gezonde spier elders uit het lichaam naar het beschadigde gebied. Er wordt dus een spierdefect gemaakt om een ander deels te herstellen, wat allesbehalve een ideale oplossing is.

Kunnen we al spieren maken in het lab? Ja!

Macroscopisch beeld van een in het labo gekweekte skeletspier. De rode vloeistof is een celcultuur medium om de spier(stam)cellen optimaal te ondersteunen.

Wat als we met behulp van de spierstamcellen in het labo skeletspierweefsel ‘kweken’ en dit gebruiken om die schade te herstellen? Deze vraag ligt aan de grondslag voor onderzoek naar de bouw van skeletspierweefsel. In ons labo kweken we menselijke spieren, iets wat vrij uniek is in de wereld van skeletspier weefselbouw technologie omdat veel andere onderzoekers werken met muizencellen. Het gebruik van menselijke cellen biedt ons een voorsprong wanneer we uiteindelijk met een werkende spier naar de kliniek zullen gaan. Ze is ook van onmiddellijk nut bij het verwerven van inzichten voor de mens. Om een menselijke spier te maken starten we met de spierstamcellen die aanwezig zijn in elke spier en verantwoordelijk zijn voor spiervezel aanmaak. We kunnen deze isoleren uit skeletspierweefsel en mengen deze met een natuurlijke gel. Deze gel ondersteunt de cellen en helpt ze spiervezels te vormen. Wanneer we gedurende minstens een week voldoende voedingsstoffen en groeifactoren voorzien, krijgen we op die manier een klein spierbundeltje van ongeveer 2 centimeter lang en 1 mm dik.

... maar we zijn er nog niet.

Zij die hun gymsessie al hebben geannuleerd in de hoop grote spieren te krijgen uit ons labo moeten we momenteel nog teleurstellen, want de huidige afmetingen zijn te klein in vergelijking met natuurlijke spieren. De gekweekte spieren groter maken is echter niet zo voor de hand liggend want meer cellen toevoegen betekent niet dat dit automatisch leidt tot grotere spieren. De voornaamste hinderpaal is de afwezigheid van bloedvaten in de gekweekte spier. Deze zijn nodig om alle cellen in de spier te voorzien van zuurstof en voedingsstoffen en om afvalstoffen af te voeren. Daarom onderzoeken we hoe bloedvaten kunnen aangemaakt worden in de kweekspier. Dit doen we door, naast spierstamcellen, ook cellen toe te voegen die de binnenste cellaag van bloedvaten vormen. Dankzij deze techniek slagen we erin om een kweekspier te maken met daarin een beginnend bloedvatennetwerk. De volgende stap is om deze netwerken verder te laten ‘rijpen’ zodat ze in staat zijn om hun functie volledig uit te voeren. Stof voor nog verschillende jaren onderzoek dus.

Steun gezocht

Een andere aanpak die onderzocht wordt is het gebruik van biomaterialen als ondersteunende draagstructuren voor de spiervezels. De meeste kweekspieren die vandaag worden gemaakt, in ons en in andere labo’s, worden ondersteund door een zachte gel, wat ze te weinig stevigheid geeft. Het idee is dat draagstructuren tijdelijk kunnen compenseren voor de gebrekkige stevigheid die de huidige constructen hebben. Zo’n draagstructuur moet echter aan heel wat vereisten voldoen. Zo moet ze stevig zijn maar toch volledig afbreekbaar want ze moet met de tijd volledig verdwijnen uit het lichaam. Het doel is namelijk dat de kweekspier niet meer te onderscheiden is van een gewone spier. Verder moet de draagstructuur biocompatibel zijn, dat betekent dat ze een optimale leefomgeving moet vormen voor de cellen die worden gebruikt om de kweekspier te maken. Dit is een cruciaal gegeven want eerdere experimenten met polymere draagstructuren moesten gestopt worden door een gebrek aan bio-compatibiliteit, de toegevoegde cellen overleefden niet. Ook moeten de mechanische eigenschappen ervan zo goed mogelijk deze van skeletspieren nabootsen. Elasticiteit is een belangrijke eigenschap van dynamische weefsels zoals spieren en pezen. Dergelijke elasticiteit kan voorzien worden door een groep polymeren genaamd elastomeren. Recent hebben we voor een aantal elastomeren getest hoe goed de menselijke spierstamcellen kunnen aanhechten, overleven, zich delen en spiervezels vormen. Binnen de groep van de elastomeren konden we er tot nu toe verschillende vinden die optimale cel overleving en vermenigvuldiging toelieten. We gaan verder op dit veelbelovende pad om een goede draagstructuur voor de cellen te vinden.

Een lijst vol toepassingen

Kweekspieren kunnen mogelijks in de toekomst dienen voor spierherstel zoals we hierboven al vermeldden, maar daarnaast zijn er nog andere veelbelovende toepassingen. Zo kan een menselijke kweekspier dienen als ziektemodel, om bijvoorbeeld onderzoek te doen naar spierziektes. Daarnaast kan het ook dienen als model om nieuwe geneesmiddelen te testen. Deze twee toepassingen kunnen ook gecombineerd worden en beiden laten toe om het gebruik van proefdieren te verminderen. Het kan ook dienen als een nieuwe manier om sportfysiologie te evalueren, door na te gaan wat het effect is van bepaalde supplementen en trainingsschema’s op de spieren. Tot slot is er de laatste tijd heel wat media aandacht voor het gebruik van dierlijke kweekspieren als alternatieve bron van vlees. Kortom, met ons onderzoek naar gekweekte spieren verwachten we vooruitgang te boeken die impact heeft op verschillende domeinen.

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Universiteit of Hogeschool
Master in de biomedische wetenschappen afstudeerrichting Biomedisch basis- en translationeel onderzoek
Publicatiejaar
2018
Promotor(en)
Prof. Lieven Thorrez
Kernwoorden