Microfluidic titer plate for stratified 3D cell culture

Abstract

Human tissues and organs are inherently heterogeneous. Their functionality is determined by the interplay between different cell types, their secondary architecture, vascular system and gradients of signaling molecules and metabolites. Here we propose a stratified 3D cell culture platform, in which adjacent lanes of gels and liquids are patterned by phaseguides to capture this tissue heterogeneity. We demonstrate 3D cell culture of HepG2 hepatocytes under continuous perfusion, a rifampicin toxicity assay and co-culture with fibroblasts. 4T1 breast cancer cells are used to demonstrate invasion and aggregation models. The platform is incorporated in a microtiter plate format that renders it fully compatible with automation and high-content screening equipment. The extended functionality, ease of handling and full compatibility to standard equipment is an important step towards adoption of Organ-on-a-Chip technology for screening in an industrial setting.

On-chip three-dimensional cell culture in phaseguides improves hepatocyte functions in vitro

“…Although the principal suitability of hepatocyte cultivation in the OrganoPlate was demonstrated, 15 a profound basic characterization of hepatocytes cultured in this biochip is still lacking. In order to make up for this and pave the way for further studies with clinical importance, the present study aimed to assess hepatocytes' behavior considering multiple performance criteria: cellular morphology and differentiation over two weeks (clustering, bile canaliculi formation, and viability), different metabolic endpoints (albumin, urea, and Cyp1A2 activity), and one clinically important toxicity assay (acetaminophen, APAP).…”

“…14 A microfluidic device with phaseguides called OrganoPlate TM was recently developed by MIMETAS and Leiden University (Leiden, The Netherlands) and first examples of cell culturing were published by Trietsch et al 15 Briefly, the cell culture chip is a commercial 364-well plate where microfluidic structures are embedded on the bottom resulting in 40 culture chambers. Each culture chamber consists of three lanes separated by phaseguides and allows the generation of a continuous passive perfusion without connection to an extra pump or tubing line.…”

Membrane-free culture and real-time barrier integrity assessment of perfused intestinal epithelium tubes

“…Figure 1 shows the OrganoPlate platform, which encompasses 40 microfluidic cell culture structures embedded in a standard 384-well microtiter plate format (Fig. 1a, b) 9,10 . Each microfluidic channel structure is comprised of three lanes that are connected to corresponding wells of a microtiter plate that function as inlets and outlets to access the microfluidic culture.…”

96-Well Format-Based Microfluidic Platform for Parallel Interconnection of Multiple Multicellular Spheroids

“…There have been efforts in developing 96-well format-based cell culturing or drug testing systems that can be operated through pipetting. [19][20][21] In this article, we present a microfluidic platform, which is compatible with 96-well automation tools and allows for parallel culturing of spherical MTs of different cell types. Its basic concept relies on our previous work 22 and includes the reliable formation of spheroids outside the platform and their subsequent direct transfer into the microfluidic platform.…”