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.
For detailed protocol of the study, please download the full Application Note.
Image text: Nanomedicine Research Group of Institute of Chemical Technology, Mumbai, India