Laser direct-write based fabrication of a spatially-defined, biomimetic construct as a potential model for breast cancer cell invasion into adipose tissue

Abstract

Epithelial-adipose interaction is an integral step in breast cancer cell invasion and progression towards lethal metastatic disease. Understanding the physiological contribution of obesity, a major contributor to breast cancer risk and negative prognosis in post-menopausal patients, on cancer cell invasion requires detailed co-culture constructs that reflect mammary microarchitecture. Using laser direct-write, a laser-based CAD/CAM bioprinting technique, we have demonstrated the ability to construct breast cancer cell-laden hydrogel microbeads into spatially defined patterns in hydrogel matrices containing differentiated adipocytes. Z-stack imaging confirmed the three-dimensional nature of the constructs, as well as incorporation of cancer cell-laden microbeads into the adipose matrix. Preliminary data was gathered to support the construct as a potential model for breast cancer cell invasion into adipose tissue. MCF-7 and MDA-MB-231 breast cancer cell invasion was tracked over 2 weeks in an optically transparent hydrogel scaffold in the presence of differentiated adipocytes obtained from normal weight or obese patient tissue. Our model successfully integrates adipocytes and gives us the potential to study cellular and tissue-level interactions towards the early detection of cancer cell invasion into adipose tissue.

Bioprinting on Live Tissue for Investigating Cancer Cell Dynamics

“…For accurate deposition of cell bioinks, the droplet-on-demand (DOD) dispensing mechanism systems are generally preferred to produce finer features with better resolution than others based on extrusion methods. 34 The main DOD bioprinting approaches include laserinduced forward transfer (LIFT) [35][36][37] and inkjetting. 20,22,38 While LIFT enables the deposition of highly viscous inks given its orifice-free nature, the droplet formation process is generally difficult to be controlled precisely.…”

Human Adipose-Derived Stromal/Stem Cell Culture and Analysis Methods for Adipose Tissue Modeling In Vitro: A Systematic Review

“…Finally, none of the screened articles exclusively studied 2D culture; therefore, all articles included in this analysis report 3D culture methods. Data and analysis hereafter exclusively refer to the 27 articles [4,5,[8][9][10][11][12][13][15][16][17][19][20][21][22][23][24][25][26]28,30,31,33,38,44,45,48] identified to be most relevant to the central goal of this systematic review. Out of the 27 relevant articles, 26 articles used adipogenic medium to differentiate hASC into adipocytes in vitro.…”

“…One critical aspect of 3D stem cell culture is developing an in vitro environment that closely mimics native in vivo conditions [11,21,25]. A widely used strategy is to seed stem cells into a scaffold that is designed to recapitulate the native extracellular matrix (ECM).…”

“…Only one article (5%) used an entirely synthetic scaffold, while the remaining articles incorporated biologically derived materials. Gelatin was identified as one of the most commonly used scaffold materials as it naturally contains RGD peptides found in many ECM components that promote cell adhesion [13,16,17,21]. Collagen is another commonly used scaffold material as it is the most abundant protein in human adipose tissue ECM and, therefore, closely recapitulates the native tissue environment [10,31].…”

Cancer Cell Direct Bioprinting: A Focused Review

“…Heinrich et al applied the extrusion technology to study the cell interactions and possible therapeutic uses in glioblastoma cancer research [ 79 ]. Other authors, such as Vinson et al, used a laser printing technology called laser direct writing (LDW) to study cancer invasion in adipose tissues [ 80 ]. Stereolithography has been used by Chen et al to evaluate the capture efficiency of cancer circulating tumor cells (CTC), which could be applied as biomarkers for early detection of cancer [ 81 , 82 ].…”