Fibroblasts And Mouse's Breast Cancer Cells Can Form Cellular Aggregates in Improved Soft Agar Culture Medium

Abstract Purpose Cancer-associated fibroblasts (CAFs) promote cancer cell growth, invasion and migration. In this research, we aimed to build cellular aggregates of mouse’s breast cancer cells (TS/A) and normal fibroblasts (LX-2) or CAFs (ME-iLX-2) in soft agar culture medium, verifying this co-culture model’s value in screening of anticancer drugs and preliminarily demonstrating the effect of CD44 in aggregates formation. Methods We improved soft agar culture medium to co-culture CAFs (NFs) and TS/A to build the cellular aggregates model, and compared the amount and area of different co-culture groups. To verify its value in screening of anticancer drugs, eugenol which has been demonstrated its anticancer effect was added into this model. The transcription of human CD44 was analyzed through Real-time quantitative PCR (RT-qPCR). Result TS/A-fibroblast cellular aggregates were formed in improved soft agar culture medium. What’s more, the amount and area of aggregates in TS/A-ME-iLX-2 co-culture group were significantly higher than in TS/A-LX-2 co-culture group. In co-culture group with eugenol, the amount and area of TS/A-ME-iLX-2 aggregates were lower than control group. The result of RT-qPCR showed that the transcription volume of human CD44 in TS/A-ME-iLX-2 aggregates was higher than in TS/A-LX-2 aggregates. Conclusion Co-culturing cellular aggregates of fibroblasts and TS/A was successfully formed in improved soft agar culture medium, and the amount and area of aggregates further confirmed CAFs’ promotion effect to cancer cells. Eugenol test showed its value in screening of anticancer drugs. RT-qPCR result demonstrated the important effect of CD44 in aggregates formation.