In vitro lung cancer multicellular tumor spheroid formation using a microfluidic device

• Published in: Biotechnology and bioengineering Vol. 116; no. 11; pp. 3041 - 3052
• Main Authors: Lee, Sang Woo, Hong, Soyoung, Jung, Boyoung, Jeong, Soo Yeon, Byeon, Jae Hee, Jeong, Gi Seok, Choi, Jaesoon, Hwang, Changmo
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.11.2019
• Subjects: Biotechnology; Cancer; Cell culture; Cell proliferation; Collagen; Collagen (type I); Continuous flow; Dosage; Drug screening; Endothelial cells; extracellular matrix (ECM); Flow velocity; Inhibitors; Kinases; Lung cancer; matrix metalloproteinase (MMP); Metastases; microfluidic device; Microfluidic devices; Microfluidics; multicellular tumor spheroid (MCTS); Perfusion; Shrinkage; Stability; Sustainability; tumor microenvironment (TME); Tumorigenesis; Tumors; lung cancer; extracellular matrix (ECM); microfluidic device; tumor microenvironment (TME); multicellular tumor spheroid (MCTS); matrix metalloproteinase (MMP); tumorigenesis
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.27114

The purpose of this study was to demonstrate self‐organizing in vitro multicellular tumor spheroid (MCTS) formation in a microfluidic system and to observe the behavior of MCTSs under controlled microenvironment. The employed microfluidic system was designed for simple and effective formation of MCTSs by generating nutrient and oxygen gradients. The MCTSs were composed of cancer cells, vascular endothelial cells, and type I collagen matrix to mimic the in vivo tumor microenvironment (TME). Cell culture medium was perfused to the microfluidic device loaded with MCTSs by a passive fluidic pump at a constant flow rate. The dose response to an MMPs inhibitor was investigated to demonstrate the effects of biochemical substances. The result of long‐term stability of MCTSs revealed that continuous perfusion of cell culture medium is one of the major factors for the successful MCTS formation. A continuous flow of cell culture medium in the in vitro TME greatly affected both the proliferation of cancer cells in the micro‐wells and the sustainability of the endothelial cell‐layer integrity in the lumen of microfluidic channels. Addition of MMP inhibitor to the cell culture medium improved the stability of the collagen matrix by preventing the detachment and shrinkage of the collagen matrix surrounding the MCTSs. In summary, the present constant flow assisted microfluidic system is highly advantageous for long‐term observation of the MCTS generation, tumorous tissue formation process and drug responses. MCTS formation in a microfluidic system may serve as a potent tool for studying drug screening, tumorigenesis and metastasis. The effective in vitro system which clarify the process of cell‐cell interaction during the initial cancer formation and the related cellular behaviors can applied for the development of effective therapeutic strategies. Lee and coworkers tried to generate lung cancer MCTS in the microfluidic chip to supply oxygen and nutrient gradient mimicking the in vivo tumor condition. Endothelial cell‐cancer cell interaction and matrix metalloproteinase effect on MCTS formation was investigated as a model cell–cell interaction and drug effect during MCTS formation.