Chondrogenesis of bone marrow and peripheral blood derived adult human mesenchymal stem cells

Azura Mansor, Pan-Pan Chong, L. Selvaratnam, Veera Sekaran Nadarajah, T. Sara, T. Kamarul

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Abstract

INTRODUCTION: Although mesenchymalstem cells (MSCs) isolation from a number oftissue sources have been described, very fewliteratures have reported successful isolation ofadult MSCs from peripheral blood or itschondrogenic differentiation for clinicalapplications. The objective of this study is toisolate MSCs derived from both human bonemarrow and peripheral blood and to comparetheir potential to undergo chondrogenesis.METHODS: Bone marrow (BM) andperipheral blood (PB) samples were collectedinto blood collection tubes containing lithiumheparin (2-4mls). Mononuclear cells wereseparated from both types of samples usingFicoll–Paque PLUS by centrifugation andsuspended in cell culture medium before beingplated onto tissue culture flasks. Suspendedcells were subsequently removed after 5 days ofculture, and adherent cells were left to grow.Cells were sub-cultured (2–5 passages) prior tofurther cellular analyses and differentiationexperiments. Chondrogenic pellets wereharvested after 4-5 weeks in culture. To assesschondrogenesis, alcian blue and safranin-Owere used to determine whether cartilage matrixproteoglycan was expressed. Chondrogenesiswas also quantified by the amount of sulphatedglycosaminoglycan (S-GAG) productionmeasured using 1,9-dimethylmethylene blue(DMMB) assay.RESULTS: Based on our results, we were ableto establish techniques for isolation of MSCsfrom BM and PB. The presence of surfacemarker proteins CD44, CD105, CD166 and theabsence of CD34 in these cells (confirmedusing flow-cytometry) indicate the highlikelihood of successful mesenchymal stem cellisolation. Histological examination revealedsignificant cellular expressions ofproteoglycans and glycosaminoglycansindicating successful induction ofchondrogenesis in our isolated MSCs. DISCUSSION & CONCLUSIONS: In vitroinduction of chondrogenesis has beendemonstrated in both bone marrow andperipheral blood-derived MSCs using 3-dimensional scaffold producing comparablecellular expressions. MSCs which are easilyisolated (and less painfully harvested) fromperipheral blood as compared to bone marrowprovides a superior alternative source for MSCsfor future clinical application (in this case forcartilage repair).    
Original languageEnglish
Pages (from-to)64
Number of pages1
JournalEuropean Cells and Materials
Volume16
Issue numberSuppl 2
Publication statusPublished - 2008

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