Microfluidic Tumor–Vascular Model to Study Breast Cancer Cell Invasion and Intravasation

Abstract

Cancer is a major leading cause of disease-related death in the world. The severe impact of cancer can be attributed to poor understanding of the mechanisms involved in earliest steps of the metastatic cascade, specifically invasion into the surrounding stroma and intravasation into the blood capillaries. However, conducting integrated biological studies of invasion and intravasation have been challenging, within in vivo models and traditional in vitro assay, due to difficulties in establishing a precise tumor microenvironment. To that end, in this work, a novel 3D microfluidic platform comprised of concentric three-layer cell-laden hydrogels for simultaneous investigation of breast cancer cell invasion and intravasation as well as vasculature maturation influenced by tumor-vascular crosstalk is developed. It was demonstrated that the presence of spontaneously formed vasculature enhance MDA-MB-231 invasion into the 3D stroma. Following invasion, cancer cells are visualized intravasating into the outer vasculature. Additionally, invading cancer cells significantly reduce vessel diameter while increasing permeability, consistent with previous in vivo studies. Major signaling cytokines involved in tumor-vascular crosstalk that govern cancer cell invasion and intravasation are further identified. Taken together, this platform will enable unique insights of critical biological events within the metastatic cascade, with significant potential for developing efficient cancer therapeutics.

Vascularized microfluidic platforms to mimic the tumor microenvironment

“…Other proteins such as fibrin have also been used as a scaffold material. Groups have utilized fibrin by itself or as a blend with different collagen percentages for various applications including cardiac, wound healing and cancer invasion studies (Chung, Ahn, Son, Kim, & Jeon, ; Lee, Park, Ryu, & Jeon, ; Mol et al, ; Nagaraju, Truong, Mouneimne, & Nikkhah, ; Purtscher, Rothbauer, Holnthoner, Redl, & Ertl, ). However, low ultimate tensile strength of this material suggests it is not well suited for tissues of high stiffness such as breast tumor microenvironments, making collagen a more desirable scaffold material (Cummings, Gawlitta, Nerem, & Stegemann, ).…”

“…Groups have utilized subtractive and additive tissue engineering processes to form microfluidic collagen scaffolds (Bettinger, Borenstein, & Tao, ; Bhatia & Ingber, ; Peela et al, ; Tien, ). Scaffolds with complex microfluidic networks have also been formed using additive methods of combining layers of natural materials formed with lithographic techniques and have been used to investigate various behaviors such as cell‐cell interactions during angiogenesis or metastasis, extravasation of breast cancer cells and interactions with the endothelium (Bhatia & Ingber, ; Bischel, Young, Mader, & Beebe, ; Farahat et al, ; Ghousifam et al, ; Jeon et al, ; Lee et al, ; Mi et al, ; Nagaraju et al, ; Peela et al, ; Price et al, ; Soleimani et al, ; Song et al, ; Y. Ma et al, ; Zheng et al, ). While these microfluidic devices have provided insight into tumor behavior, the in vitro platforms consist of small number of cells limiting the use of biological assays such as polymerase chain reaction or enzyme‐linked immunosorbent assay, or they lack a continuous endothelium failing to recapitulate the in vivo tumor microenvironment.…”

“…Groups have utilized fibrin by itself or as a blend with different collagen percentages for various applications including cardiac, wound healing and cancer invasion studies (Chung, Ahn, Son, Kim, & Jeon, 2017;Lee, Park, Ryu, & Jeon, 2014;Mol et al, 2005;Nagaraju, Truong, Mouneimne, & Nikkhah, 2018;Purtscher, Rothbauer, Holnthoner, Redl, & Ertl, 2015).…”

“…Groups have utilized subtractive and additive tissue engineering processes to form microfluidic collagen scaffolds (Bettinger, Borenstein, & Tao, 2012;Bhatia & Ingber, 2014;Peela et al, 2017;Tien, 2014). Scaffolds with complex microfluidic networks have also been formed using additive methods of combining layers of natural materials formed with lithographic techniques and have been used to investigate various behaviors such as cell-cell interactions during angiogenesis or metastasis, extravasation of breast cancer cells and interactions with the endothelium (Bhatia & Ingber, 2014;Bischel, Young, Mader, & Beebe, 2013;Farahat et al, 2012;Ghousifam et al, 2017;Jeon et al, 2015;Lee et al, 2014;Mi et al, 2016;Nagaraju et al, 2018;Peela et al, 2017;Price et al, 2008;Soleimani et al, 2018;Song et al, 2009;Y. Ma et al, 2018;Zheng et al, 2012).…”

Recapitulating and Deciphering Tumor Microenvironment by Using 3D Printed Plastic Brick–Like Microfluidic Cell Patterning

“…[1] The tumor-stroma-ECM milieu is the back bone of cancer and is an indispensable source of critical protum origenic factors that promote tumor cells growth, invasion, and and interactions between different cells can be realized in a microfluidic apparatus with a high content manner. [19][20][21][22][23] These approaches typically generate cell compartments and then release them in a common place where the phenotypical information could be analyzed for studying cell-cell interac tion. However, current microfluidicbased cell patterning faced several challenges: 1) complex manufacturing and surface modification; 2) the tangled tubing interface and liquid control system that give rise to incompatibility with standard solution handling; and 3) lack of feasible cellretrieval capability with an enclosed device.…”