A photo-polymerized poly(Nε-acryloyl l-lysine) hydrogel for 3D culture of MCF-7 breast cancer cells

Abstract

The most common in vitro cell culture platform, standard two-dimensional (2D) monolayer cell culture, often fails to mimic the tumor microenvironment, while animal models complicate research on the effect of individual factors on cell behaviors. Both are unsatisfactory in the research of molecular mechanisms of tumor development and progression and the discovery and development of anticancer drugs. In vitro three-dimensional (3D) cell culture can partially simulate in vivo conditions and 3D-cultured cancer cells can recapture many essential features of native tumor tissues. In this study, to mimic the in vivo breast tumor microenvironment, novel reduction-responsive poly(N_ε-acryloyl L-lysine) (pLysAAm) hydrogels were synthesized by rapid photo-polymerization of N_ε-acryloyl L-lysine and using N,N′-bis(acryloyl)-(L)-cystine as a crosslinker, and their physicochemical properties were characterized systemically. The results showed that the pLysAAm hydrogels were formed within 93 s under UV irradiation and exhibited almost total elastic recovery from compressions as high as 75%. The lyophilized hydrogel samples displayed a highly porous structure with interconnected pores, had an equilibrium swelling ratio of about 20, and were degraded faster in a glutathione-containing solution than in PBS solution. The biological versatility of the pLysAAm hydrogels was demonstrated by both in vitro MCF-7 cell culture and in vivo tumor formation. Compared to cells cultured as 2D monolayers, the 3D-cultured cells presented 3D cell morphology, exhibited better cell viability, expressed higher levels of pro-angiogenic factors, and showed significantly greater migration and invasion abilities. The results from assay of tumorigenicity in nude mice and histologic analysis demonstrated the enhanced tumorigenic and angiogenic capabilities of the MCF-7 cells pre-cultured in pLysAAm hydrogels. These findings suggest that pLysAAm hydrogels may be used to bridge the gap between standard in vitro cell cultures and living tissues, aid breast cancer research, and help researchers to develop novel anticancer therapeutics.