Comparative study of liquid overlay and hanging drop techniques in MCF-7 spheroids.

Comparative study of liquid overlay and hanging drop techniques in MCF-7 spheroids.
Shermin Lak,^1,* Nasrin Motamed,²
1. university of Tehran
2. University of Tehran
Introduction: Breast cancer is the most common cause of cancer in women. It is becoming recognized that two-dimensional (2D) cell cultures do not accurately reflect the conditions in vivo. Cell-cell and cell-matrix interactions cannot be imitated via 2D cell cultures. To overcome these limitations, three-dimensional spheroid culture models have developed.3D models can simulate the function of living tissues. In this study, we indicated the generation of MCF-7 spheroids.
Methods: We used hanging drop and liquid overlay techniques for generation of MCF-7 spheroids.In liquid overlay technique, the bottom of the cell culture plate was coated with agarose in order to provide non-adherence surface. Cell suspension was loaded in to plate. In hanging drop method, cells were suspended in drops on the inverted lid of the petri dish. Drops were incubated until spheroids were formed. Wells of the 96 culture plate were layered with agarose and each drop was loaded into one well.
Results: MCF-7 spheroids were generated in both techniques. Spheroid formations occurred within 96h in Liquid overlay and 48h in hanging drop method. Volume and shape of spheroids were not same in liquid overlay technique but uniform-sized spheroids were observed in hanging drop technique.
Conclusion: Three- dimensional cell cultures mimic tumor microenvironment. Since hanging drop method can generate uniform-sized spheroids, it is more useful to form MCF-7 spheroids. This technique can be used in different studies such as cell behaviors, drug screening and gene expression.
Keywords: Three- dimensional cell culture,liquid overlay technique, hanging drop technique,MCF-7 spheroid.