A novel 3D mammalian cell perfusion-culture system in microfluidic channels

Abstract

Mammalian cells cultured on 2D surfaces in microfluidic channels are increasingly used in drug development and biological research applications. These systems would have more biological or clinical relevance if the cells exhibit 3D phenotypes similar to the cells in vivo. We have developed a microfluidic channel based system that allows cells to be perfusion-cultured in 3D by supporting them with adequate 3D cell-cell and cell-matrix interactions. The maximal cell-cell interaction was achieved by perfusion-seeding cells through an array of micropillars; and 3D cell-matrix interactions were achieved by a polyelectrolyte complex coacervation process to form a thin layer of matrix conforming to the 3D cell shapes. Carcinoma cell lines (HepG2, MCF7), primary differentiated (hepatocytes) and primary progenitor cells (bone marrow mesenchymal stem cells) were perfusion-cultured for 72 hours to 1 week in the microfluidic channel, which preserved their 3D cyto-architecture and cell-specific functions or differentiation competence. This transparent 3D microfluidic channel-based cell culture system also allows direct optical monitoring of cellular events for a wide range of applications.

Chip-based liver equivalents for toxicity testing – organotypicalness versus cost-efficient high throughput

“…2d). 62 About 4 6 10 3 cells per channel were applied, which corresponds to approximately 0.05 liver lobule. Cells were perfusion-seeded into these structures, achieving a high 3D cell-cell interaction.…”

Characterization of microfluidic human epidermal keratinocyte culture

“…The use of HEK in toxicity assays (Chou et al 2002;Didier et al 1999;Monteiro-Riviere et al 2005;Rouse et al 2006;Shvedova et al 2003) makes this cell type an attractive candidate for use in microfluidic high throughput toxicity assays. Cell culture in microfluidic systems has been demonstrated by others Delamarche et al 2005;Figallo et al 2007;Flaim et al 2005;Folch and Toner 2000;Gu et al 2004;Hui and Bhatia 2007;Hung et al 2005a, Hung et al b;Kane et al 2006;Khademhosseini et al 2004;Kim et al 2006;Leclerc et al 2006a;b;Lee et al 2006;Prokop et al 2004;Raty et al 2004;Rhee et al 2005;Sin et al 2004;Song et al 2005;Toh et al 2007;Tourovskaia et al 2005;Walker et al 2002) but keratinocytes have not yet been studied at the microscale.…”

Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics

“…We have combined the front-and side-trapping mechanisms into a microfluidic channel with dimensions of 10 4 m ͑length͒ ϫ 600 m ͑width͒ ϫ 100 m ͑height͒, in arrays of micropillars ͑dimensions 30 ϫ 50 m and a 20 m gap size͒ located in the center of the microfluidic channel to filter and trap the cells using a withdrawal flow. 31,[42][43][44][45] These trapped cells maintain good viability, 3D morphology, sufficient cell-cell and cell-matrix interactions, as well as high levels of albumin secretion and UDP-glucuronyltransferase ͑UGT͒ activity for in vitro toxicology applications.…”

“…Cells are typically trapped together as cell clusters to maintain 3D cell morphology and enhanced cellular and tissue functions. 31 Here we will review various cell trapping methods in microfluidic systems and analyze their strength and limitations.…”