Controlling spheroid size

4DCELL DEVICE

6 well plates – microwells for spheroid assembly

READ-OUTS

Spheroid diameter, shape of spheroid

STANDARD CULTURE LIMITATION

Most state-of-the art methods that are used for spheroid formation are limited by the fact that it is difficult for the scientist to keep track of individual spheroid parameters. Free floating spheroids make it nearly impossible to get solid data on individual spheroid growth. Additionally, the orientation of the spheroids is different for each observation.

4Dcell MICROWELLS FOR SPHEROID ASSEMBLY ASSAY

4Dcell microwells for spheroid assembly enable the formation of spheroids with reproducible characteristics such as size and shape. The size of the spheroids depends only on cell concentration at the moment of seeding. 

EXPERIMENTAL SET-UP

In this example, a 6-well plate with microstructured gels was used to create the spheroids. Each gel contains 91 microwells and they were seeded with cell suspensions of varying concentrations; 1k, 3k, 10k and 30k cells per microwell. For this particular experiment, mesenchymal stem cells were used.  

Images of the cells were taken 24h, 36h and 72h after seeding.

DATA ANALYSIS

Spheroid size was assessed by measuring its diameter using ImageJ built-in measuring tool. The diameters of all spheroids for each different cell concentration were measured and averaged.

Figure 1. Spheroids at different cell concentrations. Microstructured gels containing 1k, 3k, 10k and 30k cells per microwell.

Figure 2. Diameter difference between spheroids. ImageJ was used to measure the diameter of the different spheroids.

Figure 3. Relationship between spheroid diameter and number of cells per microwell.

RESULTS

This simple example illustrates how the 4DCELL gels with microwells can be used to tune the size of individual spheroids. In this example it is shown how the diameter increases when a higher concentration of cells is used for seeding.