Ink Jet printing of mammalian primary cells for tissue engineering applications
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Ink Jet printing of mammalian primary cells for tissue engineering applications Rachel Saunders, Julie Gough and Brian Derby Materials Science Centre, The University of Manchester, Grosvenor Street, Manchester, M1 7HS ABSTRACT A piezoelectric drop on demand printer has been used to print primary human osteoblast and bovine chondrocyte cells. After deposition the cells were incubated at 37 C and characterised using optical microscopy, SEM and cell viability assays. Cells showed a robust response to printing exhibiting signs of proliferation and spreading. Increasing the drop velocity results in a reduced cell survival and proliferation rates but both cell types grew to confluence after printing under all conditions studied. INTRODUCTION Free form fabrication techniques such as stereolithography, fused deposition modelling and three dimensional ink jet printing are capable of manufacturing scaffolds for use in tissue engineering. These techniques can fabricate scaffolds with the precision required to incorporate a complex interconnected internal architecture and the ability to tailor the structure to both the application and the individual [1-3]. The incorporation of cells into these scaffolds however still poses a significant problem. Current cell seeding techniques, whether static or dynamic, can result in non-uniform distribution, limited penetration depth and utilises a limited variety of cell types [4-5]. It is therefore proposed that ink jet printing be used to simultaneously deposit cells and scaffold materials. Ink jet printing offers the potential of seeding cells deep within the core of a scaffold whilst controlling their location relative to scaffold architecture, varying concentrations and cell types and maximising the use of growth factors. Xu et al have already demonstrated that cell aggregates remain viable after printing using a modified thermal ink jet printer [6]. Previous work by the author has demonstrated the viability of fibroblasts printed using a piezoelectric ink jet printer [7]. Here we report a study that begins the systematic investigation into cell viability after printing using piezoelectric ink jet technology. EXPERIMENTAL DETAILS Human primary osteoblasts were isolated from femoral heads after total hip replacement surgery. Bovine chondrocytes were isolated from metacarpalphalangeal joints obtained from a local abattoir. Cells were harvested using a standard trypsination procedure. The harvested cells were re-suspended in the required quantity of DMEM (dulbeccos modified eagles medium) containing 10% foetal bovine serum, 1% Penicillin & Streptomycin and 0.005% ascorbic acid. To ensure uniform distribution and to disrupt cell clumps the final solution was gently agitated using a pipette. Printing experiments were carried out using single nozzle tubular piezoelectric jets (Microfab Inc., Plano TX, USA). The nozzle orifice diameters used 60 micron for
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both osteoblast and chondrocyte suspensions. All suspensions were printed at a rate of 10000 droplets per second, with p
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