Characterization of the interaction between fibroblasts and tumor cells on a microfluidic co‐culture device

Abstract

Fibroblasts and tumor cells have been involved in the process of cancer development, progression and therapy. Here, we present a simple microfluidic device which enables to study the interaction between fibroblasts and tumor cells by indirect contact co-culture. The device is composed of multiple cell culture chambers which are connected by a parallel of cell migration regions, and it enables to realize different types of cells to communicate each other on the single device. In this work, human embryonic lung fibroblasts cells were observed to exhibit obvious migration towards tumor cells instead of normal epithelial cells on the co-culture device. Moreover, transdifferentiation of human embryonic lung fibroblast cells was recognized by the specific expression of a-smooth musle actin, indicating the effect of tumor cells on the behavior of fibroblasts. Furthermore, multiple types of cell co-culture can be demonstrated on the single device which enables to mimic the complicated microenviroment in vivo. The device is simple and easy to operate, which enables to realize real-time observation of cell migration after external stimulus. This microfluidic device allows for the characterization of various cellular events on a single device sequentially, faciliating the better understanding of interaction between heterotypic cells in a more complex microenvironment. Keywords:Co-culture / Microfluidic / Migration / Transdifferentiation DOI 10.1002/elps.200900776 IntroductionGrowing evidences demonstrate that tumor microenvironment including stromal cells, inflammatory cells, extracellular matrix and vessels plays an important role in the development, progression and therapy of cancer [1][2][3][4][5].Fibroblasts make up the largest number of stromal cells, which are continuously influenced by soluble factors secreted by tumor cells and their migration abilities can be enhanced by tumor cells. When fibroblasts are transdifferentiated, they become activated and were termed cancerassociated fibroblasts or myofibroblasts, which will have a significant effect on the proliferation, invasion and metastasis of tumor cells [6][7][8]. Therefore, it is of great importance to investigate the heterotypic cell interactions between tumor cells and fibroblasts. At present, co-culture is often used to investigate the interactions between heterotypic cells by using Transwell assay [9,10], in which two types of cells are separated into upper and lower chambers by a semi-permeable membrane, or by bathing cells in conditioned medium [11][12][13][14]. However, these assays are limited by the tedious manual operations which require several procedures, and they are all typical end-point assays, which are difficult to realize real-time observation. In addition, they still require large consumption of cells and reagents that are expensive in amount.Microfluidics has sparked increasing interests in cellbased biological and medical analysis, due to its miniaturization, low consumption of reagents, and capabilities to integrate various experimental op...

Indirect Carcinoma-stromal Interaction Enhances Invasion of Both Fibroblast and Tumor Cells on a Microfluidic Co-culture Device

“…Homogeneous cells of HLF-HLF and A549-A549 were used as control groups. Compared to control groups, HLF co-cultured with A549 became non-uniform, getting more protrusions stretching out largely along the direction of A549, similar to the results reported by Qin (Ma et al, 2010). Protrusions, termed pseudopods, are specialized cellular structures containing an array of different proteins (Insall, 2010).…”

“…Some researchers have compared effect of normal fibroblasts with that of CAFs on tumor cells and demonstrated that the former inhibit cancer progression (Silzle et al, 2004, Kuperwasser et al, 2004. However, other researchers found out that normal fibroblasts could induce tumor growth (Hao et al, 2013, Tripathi et al, 2012, Ma et al, 2010 and quiescent fibroblasts can get activated and may be key regulators of paracrine signaling during cancer progress (Lorusso & Rugg, 2008). Therefore, it is necessary to understand the mechanism between normal fibroblasts and tumor for personalized targeted anticancer therapies.…”

A SERS‐active microfluidic device with tunable surface plasmon resonances

“…Recent advancements in chemical analysis, biomedicine, food safety inspection, environmental or metabolite detection continue to stimulate the demand for information collection by using multiplex technologies in the manner of smaller sample consumption, shorter reaction time, higher sensitivity and lower cost [1][2][3][4]. Notably, microchip-based detectionanalysis system is considered as a promising candidate to reach this goal due to its distinct advantages such as low dosage, environmental friendliness, high safety and sensitivity.…”

Biomimetic tumor microenvironment on a microfluidic platform

“…Qin et al designed a simple microfluidic device, allowing for the multiple types of cells co-culturing in the separated microchambers, with the soluble factors diffusion through the parallel cell migration regions. 54 This study reproduced the biological process of fibroblasts recruitment in the presence of the tumor cells and the activation of fibroblasts with a high expression of specific marker (a-SMA). Furthermore, the molecular mechanism underlying this activated process of fibroblasts was elucidated by Lee et al 55 It was found that this paracrine loop mediated the activated process could be inhibited with the addition of transforming growth factor beta 1 (TGF-b1) receptor/ALK5 inhibitor into the medium of fibroblasts, which might support the target of cancer therapy to prevent activated fibroblast ( Fig.…”

“…52 Meanwhile, fibroblasts are continuously induced by the diffusive factors secreted by tumor cells, and thus can be activated with the specific expression of a-smooth muscle actin (a-SMA). 52,53 Several microfluidic devices have been developed to investigate the interaction between fibroblasts and tumor cells within microenvironment by using separated microstructure 54 or pneumatic microvalves control 55 based on microscale engineering technology. Qin et al designed a simple microfluidic device, allowing for the multiple types of cells co-culturing in the separated microchambers, with the soluble factors diffusion through the parallel cell migration regions.…”