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Creating high throughput 3D in vitro cell culture models

organoids_illustration_ 3D Cell culture3D cell cultures, such as organoids, have distinct advantages over 2D cultures for diverse cell
and tissue types as they more closely mimic the natural environments of the cell. Encapsulating single
cells in a hydrogel matrix is the first step in many downstream workflows such as high throughput
single cell screening, cell-cell or cell-pathogen interaction studies, FACS of single cell
microenvironments or even the establishment of organoids.

Hydrogel spheres for 3D cell culture on the Nadia platform

Using microfluidics to make these 3D cell culture spheres confers extremely high throughput. The Nadia platform can encapsulate 600,000 cells in individual hydrogel spheres in a single 20-minute run, rendering this process suitable for both research and industrial applications.
By altering encapsulation parameters of pressure and temperature, a variety of high viscosity temperature sensitive liquid hydrogels can be flowed into droplets, from agarose to collagen gels such as Matrigel™.

     Temperature - Dolomite Bio Stirring Speed - Dolomite Bio Pressure - Dolomite Bio Time - Dolomite Bio

Few other methodologies can quickly and reproducibly generate hundreds of thousands of single cell 3D scaffolds from diverse biocompatible polymers, all whilst maintaining single cell identity. The tremendous versatility of the Nadia and Innovate platform also allows this technique to be applied to cells of diverse sizes and morphologies . Samples that have been successfully tested range from small probes such as genomic DNA or nuclei to large cells like cardiomyocytes, which can be over 60µm in diameter.

Take a look at our Plant protoplast RNA-Seq page, to learn more about the research possibilities with larger cell sizes on the Nadia Platform.

  Single cell Biology - Dolomite Biodiagram 3D Cell culture

Once droplets are produced, they can be cooled and separated from the oil phase to produce hydrogel spheres. These hydrogel scaffolds which cells can cling to can then be perfused with a variety of culture mediums for long term single cell growth studies, allowing single colony screening techniques such as ISO-Seq (Liu et al., 2019).

Here are some useful links if you want to find out more about how the Nadia Innovate can be used to generate hydrogel droplets for single cell research:

  CTA_agarose_ encapsulation_ app note_3D Cell culture CTA_hydrogel_blog_ 3D Cell culture CTA_hydrogel-howto- 3D Cell culture