2D cell culture models are routinely used in drug screening and toxicity studies, but the results may be misleading due to limited cell-cell and cell-ECM interactions. 3D cultured cells show organizational similarities to tissue and can thus be used as a relevant model for such studies. Tejal V. Pant and Ratnesh Jain from Institute of Chemical Technology, Mumbai, India, used GrowDex for the development of a lung adenocarcinoma (A549) 3D model. Microscopic observation (Fig. 1) and simultaneous Live/Dead assay showed that the spheroids start formation by day 3 and continued to grow to a size of approximately 100 µm by day 15 remaining viable till the end of culture.
Method and conclusions
0.5 % GrowDex was prepared by diluting the stock solution with culture medium. 100 µl of diluted GrowDex was transferred to a 96-well microplate, and A549 cells were seeded on top. Spheroids were recovered from the matrix using GrowDase enzyme subsequent to overnight incubation at 37°C.
Figure 1. Cell membrane and Nucleus of A549 spheroid stained with Cell Vue Maroon (red) and DAPI (blue) as observed using Confocal microscope (A) Individual channel and merged image in 2D, and (B) 3D stack showing cross section of spheroid. Scale bar is 20 µm.
The team concluded that GrowDex provides an easy to use 3D support for development of A549 spheroids. Suitable optical properties of this matrix enable facile visualization of spheroids microscopically. The spheroids can be used for assessing toxicity of different formulations used for drug delivery. These can also be used in combination with other types of lung cells, fibroblasts and/or macrophages in co-culture studies.
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Nanomedicine Research Group of Institute of Chemical Technology, Mumbai, India