Rapid Microfluidic Formation of Uniform Patient-Derived Breast Tumor Spheroids

Abstract

Breast cancer is a highly complex, heterogeneous, and multifactorial disease that poses challenges for rapid and efficient treatment and development of personalized therapy. Here, we describe a rapid and reliable method to generate threedimensional (3D) tumor spheroids in vitro that recapitulate an individual patient's tumor for testing treatments. By employing droplet microfluidics and scaffold materials, tumor cells were encapsulated into a large number of Matrigel-in-oil droplets with precise control over cell numbers and components per droplet. After removal of the oil, large numbers of uniform tumor spheroids were formed within a few hours via Matrigel-supported cell selfassembly. Our microfluidic technique produces uniform-sized tumor spheroids in less than 1 day. This method was used to reproducibly and rapidly generate uniform-sized tumor spheroids derived from patients' breast tumor tissues. As a proof-of-concept application, this method was used to quickly evaluate cancer treatments. We demonstrated that our microfluidic patient-derived tumor cultures not only preserve the genetic characteristics of the original tumor tissue but also provide heterogeneous responses to targeted therapies within 2 days. We believe this method will enable a timely and reliable 3D in vitro culture model, which may be applicable to personalized treatment prediction, drug discovery, and toxicity testing.

Acoustic Droplet Printing Tumor Organoids for Modeling Bladder Tumor Immune Microenvironment within a Week

“…[5] Animal models and 2D cell culture models are widely used in the study of human diseases, whereas animal models are limited by the physiological differences between humans and animals, which makes it difficult to simulate the histological and pathological characteristics of patients; while 2D cell culture models cannot reflect the complexity and cellular diversity of 3D solid tumors. [6][7][8][9] Therefore, it is of great significance to construct a 3D model in vitro to reproduce the physiological characteristics of patients and realistically simulate the tumor microenvironment.…”

“…Organoid is a miniaturized organ model cultured in vitro in three dimensions, capable of recreating the internal complexity of tissues, derived from cells from patient tissues, embryonic stem cells, or induced pluripotent stem cells through selforganized 3D culture, [10][11][12] while some organoids can be constructed by various biofabrication strategies, such as 3D bioprinting methods and microfluidic chips. [2,6] Organoid models have been used in a variety of tumor biology studies. [13][14][15] Due to the highly consistent pathological characteristics of the tumor organoid and the patient's tumor, they are mainly used for personalized and precise treatments for patients, as well as for pharmacokinetic experiments before the drug enters clinical trials and researches on tumor mechanisms, cell migration, and metastatic.…”

Droplet Microfluidics for Tumor Drug‐Related Studies and Programmable Artificial Cells

“…This can enable multiple cell type co‐cultures in predefined microenvironments, that better simulate in vivo tissue complexity, thus making possible the development of high throughput platforms for drug treatments tailored to patients (Figure 2d ). [ 64 ]…”

Parallel and large‐scale antitumor investigation using stable chemical gradient and heterotypic three‐dimensional tumor coculture in a multi‐layered microfluidic device

“…[12,13] Furthermore, microfluidic integration on a large-scale enables highthroughput operation and investigation of cell/tumor samples. [14,15] In the past several decades, research scholars around the globe have developed many microfluidic tumor manipulation systems that depend on passive microwells, [16][17][18][19] microdroplets, [20,21] microhydrogels, [22][23][24] active pneumatic microstructures (PμSs), [25][26][27] as well as 3D acoustic tweezers [28,29] for controllable 3D tumor cultivation. These advances have provided several beneficial properties, such as facile and efficient operation, homogeneous and large-scale tumor production, and throughput monitoring.…”

“…These advances have provided several beneficial properties, such as facile and efficient operation, homogeneous and large-scale tumor production, and throughput monitoring. In addition, microfluidic tumor platforms have been widely applied for exploration of the tumor microenvironment, [16,24,30,31] tumor progression, [21,32] chemotherapy, [18][19][20]27,28,33] imaging, [25,34] and tumor-specific molecule detection. [35] However, microfluidic progress enabling the real-time detection and synchronous evaluation of 3D tumor responses to drugs at various concentrations by using a single device, was limited.…”