Novel egg white based 3D cell culture system for breast cancer research.

Abstract

Abstract Abstract #4060 Introduction: Over the last few years, much attention has been paid to the development and utilization of three-dimensional (3D) cell culture to replace classical two-dimensional (2D) monolayer cell culture systems grown as a flat layer on plastic. Not completely surprisingly, there are tremendous differences between cells in 2D and 3D culture. However, currently available 3D cell culture media are expensive for regular use by the majority of research laboratories and thus large scale use of 3D cell culture system is presently cost prohibited. In our search for a reliable and economically viable replacement for the expensive commercial 3D media, we hypothesized that avian egg white could be a potential alternative.
 Methods: Egg white-based chamber slide: Separated egg white from the chicken eggs and coated each wells of an eight well culture chamber with 80μl of egg white by careful heating at 600C on a heating block. After 30-60 minutes, the egg white becomes a semi-solid that adheres to the bottom of the well. The unstuck egg white is washed out by adding 500µl of growth medium and removing it slowly using a Pasteur pipette.
 Cell culture and analysis: To each well 2x103 immortalized human breast epithelial cells (MCF10A), were added in 0.5ml of appropriate culture medium. We have analyzed the growth curve of the acini, lumen formation, apoptosis and cell proliferation at different days post-culture. Apico/basal polarization of the acini was analyzed using appropriate antibodies. We have also cultured IGFR1-transformed MCF10A cells, different established human cell lines (MCF7, HEK293, HeLa, LNCaP, and Saos-2), and MMTV-PyMT transformed mouse mammary epithelial cells in their appropriate growth media. For comparative studies, we used a well-established reconstituted basement membrane matrix preparation (BM).
 Results: Our analysis shows that this simple avian egg white based system supports growth of cells in 3D, with significantly decreased cost. Specifically, the growth of MCF10A in egg white-based medium results in formation of acini with hollow lumens, apoptotic clearance of the cells in the lumen, and apico/basal polarization comparable to what has been described using established 3D culture media. There was no significant difference in MCF10A proliferation and acini size between egg white and BM. We have also observed similar morphology for different established cell lines, oncogene-transformed MCF10A, and mouse mammary epithelial cells between egg white and BM.
 Conclusion: Our data convincingly argue that egg white can be used as a suitable alternative model for 3D cell culture studies. We strongly believe that this simple and inexpensive method should allow researchers to perform 3D cell culture on a regular basis, and thus result in a dramatic increase of utilization of the 3D cell culture in research. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 4060.