3D Cell Cultures

Hydrogels are becoming more and more popular as platforms for three-dimensional (3D) cell culture.

3D hydrogel matrices have been used for a variety of applications, including tissue engineering of micro-organ systems, drug delivery, cytotoxicity testing, and drug screening.

Moreover, 3D cell culture is applied for investigating cellular physiology, stem cell differentiation, tumor models and for studying interaction mechanisms between cells and extracellular matrix.


Miniaturization and multiplexing of cell culture on biochip is attracting a growing interest with the development of single-cell assays and cell-based biosensors.

In a research performed at Laboratoire de Génie Enzymatique, Membranes Biomimétiques et Assemblages Supramoléculaires (GEMBAS) together with Institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS) at Université Claude Bernard Lyon 1, collaborators reported rapid methods for microwell preparation and controlled dispensing of different cell lines within these fabricated microwells.

HeLa cells were encapsulated in alginate beads suspended in DMEM solution and visualized through a glass coverslip.

The alginate beads were generated using sciDROP PICO technology mounted on a sciFLEXARRAYER S3.

 

 

 

Co-culture strategies are foundational in cell biology. These systems, which serve as mimics of in vivo tissue niches, are typically poorly defined in terms of cell ratios, local cues and supportive cell–cell interactions. In the stem cell niche, the ability to screen cell–cell interactions and identify local supportive microenvironments has a broad range of applications in transplantation, tissue engineering and wound healing.

 

A microfluidic platform for the high-throughput generation of hydrogel microbeads for cell co-culture was introduced. Encapsulation of different cell populations in microgels was achieved by introducing in a microfluidic device two streams of distinct cell suspensions, emulsifying the mixed suspension, and gelling the precursor droplets.


HTS of protein-protein interactions in mammalian cells using transfected cell arrays

Investigation of protein-protein interactions (PPI) within cells is essential for the elucidation of biological processes and cellular networks.

The two-hybrid system is the most commonly used method for PPI analysis.

A high-throughput, cost effective method for analysing PPI directly in mammalian cells was established: the cell array-based protein-protein interaction assay (CAPPIA).


Immobilization and activity of bacteria for prokaryotic chips

Immobilization of bacteria is an important step in the development of prokaryotic cell chips. The sciFLEXARRAYER S3 has been used to spot bacteria in nanoliter volumes. The activity of the bacteria has been tested.

In most cases, the bacteria maintained their activities after 24 hours. This indicates that they also survived the spotting process when comparing the luminescence activities before and after spotting.


Drug screening and siRNA transfection using a “DropChip” cell array

In order to perform information-rich high-throughput screening, a “DropChip” microarray has been developed for multiplexed cell-based assays.

With the arrayed cell culture nanoliter droplets, synergic effects of siRNA and cisplatin were analysed. With up to 100 cells per drop, cell behavior at the individual cell level could be analysed, using high resolution fluorescence microscopy and automated image analysis.

This novel cell array format could enable highly informative functional genomic studies and large scale in vitro toxicity testing.


Multiplexed Toxicity Assays

The development of high-throughput and highly relevant in vitro toxicity assays can help to limit the extent of animal testing.

An in-vitro method for toxicity testing has been established, combining bioassay and morphometric endpoints.