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Cytomatrix 
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The Use of a 3D Matrix to Grow Replicate Aliquots of Adherent Cells for Assays and Microscopy |
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Erika von Schild, Leslie Arnold, John T. Flickinger and Todd Upton
Cytomatrix, Woburn MA, 01801 |
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Current methodologies for growing replicate samples of adherent cells have existed in cell-culture for many years. However, this two-dimensional culture methodology has proven inadequate for addressing many of the more complex questions regarding how cells interact in the more complex environments that exist in vivo. This is evidenced by the poor success rate of in vivo drug efficacy despite their successful performance in high throughput screening assays. It is predicted that an assay system that more closely resembles the in vivo growth environment of cells (i.e. 3D cell growth) will be more reliable in predicting drug efficacy in vivo. We tested whether we could generate multiple similar replicates of adherent cells grown in 3D, which could be further used in downstream assays. To evaluate this we used the Cytomatrix™ Spinner System, a cell culture device that combines a three-dimensional growth substrate, Cytomatrix™, and traditional spinner flask technology. Cells (both primary cells and cell lines) were seeded onto multiple individual Cytomatrix™ units within a specially designed holder. The holder, with seeded matrix units, was then placed in a spinner flask and the cells were cultured under conditions of convective flow. At various time points, individual matrices were removed from the culture and analyzed using a number of traditional cell culture assays. When we examined the Cytomatrix™ units for confluence or cell number we found that there were no differences, regardless of their position within their own holder or when compared to units in additional holders within the same vessel. These pieces could be subsequently used in immunofluoresence, co-culture, microscopy and secretion studies. These results demonstrate that by using the described culture system, it is possible to obtain true aliquots of adherent cells. |
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