The effect of growth and differentiation factor 5 in tenogenic differentiation

T. Kamarul, S-L. Tan, Lakshmi Selvaratnam, C-C Tai

Research output: Contribution to journalMeeting Abstractpeer-review

2 Citations (Scopus)

Abstract

INTRODUCTION: Growth and differentiationfactor 5 (GDF-5 or bone morphogeneticproteins 14) is a multifunctional proteinmolecule that plays an important role in tissuedifferentiation 1. Although its role in tendonrepair has been described, it use in vitrotenogenic differentiation from MSC has notbeen determined. Owing to the limitation ingetting allograft for tendon repair,mesenchymal stem cell provides an alternativecell based therapy for damaged tendons. Wetherefore conducted an in vitro trial to isolate,characterize, proliferate and differentiate rabbitbone marrow derived MSCs with GDF-5 inview of future in vivo applications. METHODS: Rabbit MSCs isolated from rabbitbone marrow via gradient centrifugationmethods and expanded using cell culturetechniques were used in this study. Total RNAwere extracted from MSCs and further analyzedusing our custom designed primers based onsequence information attained from GenBankdatabase. Concurrently, MSCs fixed andembedded in epon were viewed usingtransmision electron microsope (TEM). Tostudy the possibility of tenogenicdifferentiation, MSCs seeded in six-well cultureplates were maintained in culture mediumsupplemented with GDF-5 at differentconcentrations for 4 days. The collagenexpression of the MSCs was determined usingSircolTM collagen assay. Real-time PCR wasconducted to assessed the gene expressionprofile of the type-I collagen and Scleraxis inMSC in growth medium, differentiationmedium with and without GDF-5, and tenocyte.RESULTS: Plastic adherent mononuclear cellsappear heterogeneous with the majority of cellspossessing fibroblastic morphology. Themononuclear cells isolated expressed CD29 andCD166 proteins but were negative for CD 45,which is similar to that reported previously.TEM images demonstrated ultra structureswhich may be of interest for further biologicalunderstanding. Collagen expression in MSCappears to up-regulated with increasing GDF-5 concentrations (table 1). MSC grown intenogenic medium showed an increase in theirgene expression in type-I collagen andScleraxis as compared to undifferentiated MSCand autologous cultured tenocytes.DISCUSSION & CONCLUSIONS: Ourtechnique of MSC isolation appears to producereplicable results validated using ourcharacterization methods. Previous studyreported that GDF-5 plays an important role intendon repair1,2. In our study, tenogenicdifferentiation appears achievable and is usefulfor future clinical application namely in tendonrepair using differentiated MSCs.  
Original languageEnglish
Pages (from-to)47
Number of pages1
JournalEuropean Cells and Materials
Volume18
Issue numberSuppl 1
Publication statusPublished - 2009

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