Development of three dimensional culture systems for embryonic stem cells

This meeting abstract discusses the development of three dimensional culture systems for embryonic stem cells. The capacity of pluripotent, self-renewing embryonic stem (ES) cells to differentiate into any cell type of the adult body holds great potential for their use in tissue engineering and cellular therapy.
However, current two dimensional (2D) culture techniques are hindering their large scale culture and efficient differentiation. Three dimensional (3D) scaffolds can mimic the in vivo micro-environment of ES cells, leading to sustained pluripotency and enhanced differentiation into mature cell phenotypes. We are focusing on the development and application of electrospun poly(lactic-co-glycolic acid) (PLGA) scaffolds and self assembling Fluorenylmethoxycarbonyl (Fmoc) peptide hydrogels to the culture of ES cells. Parameters for electrospinning 20% PLGA in Hexafluoroisopropanol (HFIP) have been established. The resulting non woven meshes were comprised of fibres rang-
ing between 0.2 and 1.5 mm in diameter. Mouse ES cells cultured on the surface of RGD functionalised dipeptide hydrogels formed colonies reminiscent of those observed in standard 2D culture conditions. Cells seeded within the gels remained viable after 24hours. Future work will focus on assessing the survival of ES cells within the Fmoc-dipeptide hydrogels over a longer time period, and the successful application of electrospun PLGA to ES cell culture. Long term aims include modification of both 3D culture environments with oligosaccharides or extracellular matrix proteins in order to better replicate the environment stem cells experience in vivo.