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Presentation Abstract

Title: CO-CULTURES OF ENDOTHELIAL PROGENITOR CELLS ANDMESENCHYMAL STEM CELLS FOR THE GENERATION OFVASCULARISED TISSUE ENGINEERED GRAFTS

Category: Stem Cells and Tissue Engineering

SecondaryCategory: Endothelial Cells/Hemangioblasts

PresentationStart: 6/13/2013 6:00:00 PM

PresentationEnd: 6/13/2013 8:00:00 PM

PosterBoardNumber:

T-1193

AuthorBlock

Mark Chong1, Dedy Sandikin1, Renyi Teo2, Wenhao Leow2, Junwei Goh2,Toon Tien Foo2, Mahesh Choolani1, Jerry K Y Chan31Obstetrics and Gynaecology, National Univ of Singapore, Singapore, Singapore,2Centre for Biomedical and Life Sciences, Singapore Polytechnic, Singapore,Singapore, 3Reproductive Medicine, KK Women's and Children's Hospital,Singapore, Singapore

Abstract: Engineered bone tissues are currently limited by inadequate vascularisation invivo following implantation. Recent research has turned to the use of angiogeniccell sources, including endothelial progenitor cells (EPCs) to generate pre-vascularised tissue prior to implantation. In this study, co-cultures of umbilicalcord-blood derived EPCs and fetal bone marrow mesenchymal stem cells (MSCs)were studied for use in the pre-vascularisation of tissue engineered constructs.Cells were fluorescently-labelled to facilitate imaging and identification in co-cultures. Culture conditions were then optimised in monolayer cultures, andsubsequently extended to three-dimensional cultures for applications in tissueengineering. In monolayer cultures, time-lapsed observations demonstrate thecocultures to generate networks, following endothelial cell aggregation andangiogenic sprouting, in a process akin to that of physiological vaculogenesis.

10/25/2015 Abstract Print View

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Cultures of EPC in MSC-conditioned media failed to elicit similar results,suggesting the need for direct cell contact and the stromal/supportive role of MSCin the system. A modified image analysis method was then developed tocharacterise vasculogenic events, and used in the optimisation of cultureconditions to elicit maximal pre-vascularisation. It was established that culture incomplete endothelial growth medium extensive prevascular networks (1.5-foldincrease in tube length over conventional culture conditions, p<0.01). In addition,MSC were critical for the provision of stromal support, extending vascularlongevity in a dose-dependent manner. Extending these results to three-dimensional conditions relevant for tissue engineering, the EPC:MSC co-cultureswere induced to form spheroids by culture on non-adhesive plates. Spheroidsmeasuring 500 um in diameter were generated with well-established endothelialnetworks, which were sustained for up to 21 days of culture. Scaling up tophysiologically relevant conditions, co-cultured spheroids were loaded into tissueengineering scaffolds and maintained in a bioreactor for 3 weeks. Outgrowth ofcells from spheroids onto the scaffolds was observed and pre-vascularisednetworks were found to be present, as opposed to direct cell seeding methods. Inconclusion, a method to generate pre-vascularised tissue engineered constructswas developed and shown to have several promising features. Work in progressincludes evaluation of the constructs in a murine model.

The information about presentations at the ISSCR 11th Annual Meeting is available forplanning purposes only and is strictly embargoed until presentation.

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