Abstract Template 1 (Basic Science)

The Role of Transforming Growth Factor-Beta in Human Mesenchymal Stem Cells Differentiation

1Last Name, First Name; 2Mahmood, MM; 1Ching, P-K; 2Devi, M; 1Craig, R 1Department, Institute, Street Address, City, Country, 2Department of Orthopaedic Surgery, Faculty of

Medicine, University of Malaya, Lembah Pantai, Kuala Lumpur, 50603, Malaysia.E-mail of Applicant

INTRODUCTION:Mesenchymal stem cells (MSCs) have been studied extensively in order to optimize the best culture condition to differentiate them into tenogenic lineage. Transforming growth factor-beta (TGF-β) is one of the growth factors which have been used for MSCs differentiation. In this study, our objective was to determine the role of TGF-β in human MSCs during tendon regeneration and differentiation.

MATERIALS & METHODS:Ethics approval was granted by Ethics Committee (Reference number: 123.45). Human MSC were isolated from bone marrow procured from the consented patients and expanded in vitro following methods previously established1. To induce the MSCs tenogenic differentiation, MSCs were treated with TGF-β at 0, 10 and 50 ng/ml for 3 days. The total collagen expression of the MSCs was determined by using Assay X. Gene expression profile of collagen A, B, C and D were analyzed by real-time PCR analysis. Data was analyzed by using SPSS statistical software package (ver. 13.0).

RESULTS:Primary human MSCs culture showed fibroblastic morphology with a spindle-shaped appearance. When treated with TGF-β, there was an increase in total collagen expression (Figure 1). Real time PCR analysis showed different gene expression profile in tenogenic-MSCs when different amount of TGF-β were (Figure 2). At 50ng/ml of TGF-β, the genes related to tendon collagen fibres formation, Col-A and Col-B were significantly increased (p<0.05). However, the increase in Col-C expression was not observed. Tenogenic marker genes, Gyf and Tgs, and undifferentiated stem cell marker, Ips, were also increased in MSCs cultured in 50ng/ml TGF-β.

Figure 1: Legend._______________________

Figure 2: Legend. ______________________

DISCUSSIONS:The tenogenic specific genes expression (Gyf, Tgs and Ips) appeared to be highest when treated with TGF-β. While previous studies have demonstrated the importance of TGF-β in MSCs transformation, our study contributes to further observation that a minimum amount of 50ng/ml of TGF-β concentration is required to elicit optimal tenogenic differentiation in human MSCs.

CONCLUSION:The use of TGF-β in the differentiation of MSCs to tenogenic MSCs is dose dependant with an observed optimal TGF-β concentration of 10-50ng/ml required to produce an appropriate tenogenic response.

REFERENCES: 1. Tan et al., International Society for Stem Cell Research (ISSCR) 7th Annual Meeting 2009; Pg226.

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