An integrated microfluidic cell culture system for high-throughput perfusion three-dimensional cell culture-based assays: effect of cell culture model on the results of chemosensitivity assays

Abstract

Although microfluidic cell culture systems are versatile tools for cellular assays, their use has yet to set in motion an evolutionary shift away from conventional cell culture methods. This situation is mainly due to technical hurdles: the operational barriers to the end-users, the lack of compatible detection schemes capable of reading out the results of a microfluidic-based cellular assay, and the lack of fundamental data to bridge the gap between microfluidic and conventional cell culture models. To address these issues, we propose a high-throughput, perfusion, three-dimensional (3-D) microfluidic cell culture system encompassing 30 microbioreactors. This integrated system not only aims to provide a user-friendly cell culture tool for biologists to perform assays but also to enable them to obtain precise data. Its technical features include (i) integration of a heater chip based on transparent indium tin oxide glass, providing stable thermal conditions for cell culturing; (ii) a microscale 3-D culture sample loading scheme that is both efficient and precise; (iii) a non-mechanical pneumatically driven multiplex medium perfusion mechanism; and (iv) a microplate reader-compatible waste medium collector array for the subsequent high throughput bioassays. In this study, we found that the 3-D culture sample loading method provided uniform sample loading [coefficient of variation (CV): 3.2%]. In addition, the multiplex medium perfusion mechanism led to reasonably uniform (CV: 3.6-6.9%) medium pumping rates in the 30 microchannels. Moreover, we used the proposed system to perform a successful cell culture-based chemosensitivity assay. To determine the effects of cell culture models on the cellular proliferation, and the results of chemosensitivity assays, we compared our data with that obtained using three conventional cell culture models. We found that the nature of the cell culture format could lead to different evaluation outcomes. Consequently, when establishing a cell culture model for in vitro cell-based assays, it might be necessary to investigate the fundamental physiological variations of the cultured cells in different culture systems to avoid any misinterpretation of data. As a whole, we have developed an integrated microfluidic cell culture system that overcomes several technical hurdles commonly encountered in the practical application of microfluidic cell culture systems, and we have obtained fundamental information to reconcile differences found with data acquired using conventional methods.

The Study of the Frequency Effect of Dynamic Compressive Loading on Primary Articular Chondrocyte Functions Using a Microcell Culture System

“…Moreover, it was also found that the generated compressive strain decreased with the increase of operating frequency (Figure 2(b)). At a given pneumatic pressure, the dynamic process to load a pneumatic chamber with an air pressure is time-dependent [28, 29]. Higher operating frequency implies shorter imposition time (e.g., 2.0, 1.0, and 0.5 sec round −1 for the 0.5, 1.0, and 2.0 Hz, resp.)…”

Bioreactor-engineered cancer tissue-like structures mimic phenotypes, gene expression profiles and drug resistance patterns observed “in vivo”

“…In order to partially address the latter issues, the p3D culture system could also be extended to include the co-culture of a variety of tumor cells with additional, non-transformed cell types such as mesenchymal stromal cells, tumor-associated fibroblasts or endothelial and immunocompetent cells, in order to explore tumor specific microenvironmental features [4]. On the other hand, the use of tumor TLS generated in p3D, eventually produced in miniaturized systems [18,49], properly adapted for the application of direct perfusion, could help to overcome limitations inherent in the use of human cell lines xenografts for drug screening. This could be especially relevant regarding costs, time requirements and confounding effects of murine stromal and innate immune system cells.…”

Microfluidic devices for cell cultivation and proliferation

“…By using a heater chip, stable thermal conditions for cell cultivation was provided and non-mechanical pneumatically driven multiplex medium perfusion mechanism (1.6 to 40.7 ll/h) was utilised. 68 For anti-cancer drug screening, Chen et al 52 developed a microfluidic array for 3D cell culture. The device consisted of 32 flow units on a standard 96 well plate where each unit had 3 wells (flow inlet, cell culture chamber, and flow outlet).…”