Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament of Healthy Teeth

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Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament of Healthy Teeth

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Publication 2-year master student thesis
Title Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament of Healthy Teeth
Author Lagerholm, Sara
Date 2019
English abstract
ABSTRACT: Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament of Healthy Teeth AIM: To isolate and culture viable cells from the periodontal ligament and confirming their identity as mesenchymal stem cells. METHODS AND MATERIALS: Healthy premolars were collected at the time of orthodontic extractions. The middle 1/3 of the periodontal ligament was scraped and subsequent cell isolation was performed using an enzymatic method; yielding single cell isolates. Cells were cultured and maintained under standard culture conditions. Cell characterization was performed by flow cytometry using two sets of cell surface markers; one known to be present and one known to be absent in mesenchymal stem cells. Ability of the cells for in vitro differentiation into adipogenic and osteogenic lineages was tested using specifically formulated media supplements. RESULTS: Cells were successfully isolated from 11 of 13 teeth and were maintained as adherent cultures for up to 8 generations. Cellular expression of positive markers; CD73, CD90 and CD44 were confirmed by flow cytometry. For the negative marker panel, expression of CD45, CD34, CD11b, CD19 and HLA class II were not detectable. The expression of CD105 was inconclusive. As determined by phenotypic changes, cells appeared to have undergone adipogenic and osteocytic differentiation at 21 days. CONCLUSION: This study has resulted in successful isolation and partial characterization of mesenchymal stem cells from the periodontal ligament of healthy teeth. Non-invasive access to these cells, provides an excellent tool for future studies, potentially leading to beneficial knowledge transferable to the dental clinical situation.
Swedish abstract
ABSTRAKT: Isolering och karaktärisering av mesenkymala stamceller från periodontalligamentet hos friska tänder SYFTE: Att isolera och odla celler från periodontalligamentet samt karaktärisera dem som mesenkymala stamceller. MATERIAL OCH METOD: Friska premolarer gjordes tillgängliga vid ortodontiska extraktioner. Den mellersta 1/3 av periodontalligamentet skrapades varpå en enzymatisk metod användes för isolering av individuella celler. Resulterande celler odlades under standardiserade metoder. Karaktärisering av celler skedde genom flödescymetri med 2 olika paneler av cellyta markörer; en för etablerat positiva uttryck och en för kända negativa uttryck hos mesenkymala stamceller. Möjlighet av celler att differentieras in vitro till adipocyter och osteocyter testades genom tillförsel av specifika substanser till odlingsmediet. RESULTAT: Celler från 11 av 13 tänder isolerades och odlades framgångsrikt adherenta till odlingsytan i upp till 8 generationer. Celluttryck av de positiva markörerna CD73, CD90 samt CD44 bekräftades genom flödescymetri. Inget uttryck observerades för den negativa panelen CD45, CD34, CD11b, CD19 eller HLA class II. Uttrycket av CD105 kunde inte fastställas pga ofullständigt data. Försök till differentiering av celler till adipocyter och osteocyter visade på fenotypiska förändringar efter 21 dagar. SLUTSATS: Den här studien har bidragit till framgångsrik isolering och delvis karaktärisering av mesenkymala stamceller från periodontalligamentet hos friska tänder. En icke-invasiv metod av detta slag, resulterande i tillgång till denna cellpopulation utgör ett lovande verktyg för framtida studier med goda möjligheter till ytterligare kunskap applicerbart till kliniska situationer inom tandvården.
Publisher Malmö universitet/Odontologiska fakulteten
Language eng (iso)
Subject Mesenchymal stem cells
Osteogenic differentiation
Adipogenic differentiation
Extracted teeth
Periodontal ligament
Handle http://hdl.handle.net/2043/29718 Permalink to this page
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