Printing cancer cells into intact microvascular networks: a model for investigating cancer cell dynamics during angiogenesis

Abstract

While cancer cell invasion and metastasis is dependent on cancer cell-stroma, cancer cell-blood vessel, and cancer cell-lymphatic vessel interactions, our understanding of these interactions remain largely unknown. A need exists for physiologically-relevant models that more closely mimic the complexity of cancer cell dynamics in a real tissue environment. The objective of this study was to combine laser-based cell printing and tissue culture methods to create a novel ex vivo model in which cancer cell dynamics can be tracked during angiogenesis in an intact microvascular network. Laser direct-write (LDW) was utilized to reproducibly deposit breast cancer cells (MDA-MB-231 and MCF-7) and fibroblasts into spatially-defined patterns on cultured rat mesenteric tissues. In addition, heterogeneous patterns containing co-printed MDA-MB-231/fibroblasts or MDA-MB-231/MCF-7 cells were generated for fibroblast-directed and collective cell invasion models. Printed cells remained viable and the cells retained the ability to proliferate in serum-rich media conditions. Over a culture period of five days, time-lapse imaging confirmed fibroblast and MDA-MB-231 cell migration within the microvascular networks. Confocal microscopy indicated that printed MDA-MB-231 cells infiltrated the tissue thickness and were capable of interacting with endothelial cells. Angiogenic network growth in tissue areas containing printed cancer cells was characterized by significantly increased capillary sprouting compared to control tissue areas containing no printed cells. Our results establish an innovative ex vivo experimental platform that enables time-lapse evaluation of cancer cell dynamics during angiogenesis within a real microvascular network scenario.

Clinical study of contrast-enhanced digital mammography and the evaluation of blood and lymphatic microvessel density

“…Tumours with abundant stroma and collagen, so called desmoplastic, tend to collapse the capillaries and have shown a lower perfusion than tumours with a lax stroma. [22][23][24][25] The microenvironment is a complex histopathological structure composed mainly by a number of extracellular matrix molecules, macrophages, myoepithelial and endothelial cells and is increasingly recognized as a major regulator of carcinogenesis. Presently, there is no way to quantify the abnormality of the capillary network in a tumour and its microenvironment; thus, estimating the number of vessels is a rough estimation of the proliferation of blood and lymph capillaries that indicates the malignant nature of the tumour mass.…”

Laser Direct‐Write Onto Live Tissues: A Novel Model for Studying Cancer Cell Migration

“…We do know that most cells remain viable including CD11b positive macrophages and other interstitial cells, yet further characterization is needed to identify the various cell populations. Moreover, we have yet to confirm cancer cell viability past 5 days in culture with rat mesenteric tissues (Phamduy et al, ). Additional investigation is needed to define the temporal resolution and maximum duration of experiments.…”

Current developments in 3D bioprinting for tissue engineering

“…Current two-and three-dimensional models present limitations as the complexity of tumours are not replicated and the tissues do not possess vascular networks [49]. Using Bioprinting techniques, complex interactions such as cancer cell dynamics during vascularization can be researched [50,51]. Metastasis of cancer cells is one of the biggest dangers of tumours.…”