New well plate differentiates itself from the crowd

The AggreWell 400 controls the formation of the three-dimensional cell aggregates known as embroid bodies (EBs) that are crucial to many of the protocols used when inducing human embryonic stem cells (hESCs) to differentiate.

Conventional methods of forming EBs include scraping undifferentiated cell cultures to release large, poorly defined clumps of cells or suspending single cell hESC populations in hanging drops and placing the resulting aggregates in suspension culture.

Both of these methods form EBs that are heterogeneous in both size and shape and lead to inefficient and uncontrollable differentiation.

In contrast, the AggreWell plates contain microwells that that aggregate the hESCs into highly uniform EBs that make differentiation experiments more reproducible.

According to research published in the journal Stem Cells​ by a team of researchers led by Professor Peter Zandstra from the University of Toronto, the direction in which cells differentiate is affected by three factors: the composition of the hESCs, the size of the colonies and the size of the EBs.

“The AggreWell 400 plate standardizes a critical step in the development of embryonic and induced pluripotent stem cells into clinically-relevant mature cells,”​ said Dr Clive Glover, Stemcell Technologies’ Product Manager for Pluripotent Stem Cell Biology.

“These cells have been difficult to work with because of the lack of standardized methods to culture and differentiate them. The AggreWell 400 plate is the latest product in Stemcell Technologies’ portfolio that addresses the need for standardszed methods and tools for stem cell research.” ​

Typically a single cell suspension will be added to a plate before centrifuging and then culturing for 24 hours.

The EBS can then be harvested by simply pipetting the contents of a well into a 40uM cell strainer which collects the EBs and allows the single cells to be washed away.

According to the company, because the number of microwells in each of the AggreWell plates is known, it is easy to control the size of the resulting EBs simply by adjusting the number of cells added per well.

The more labour-intensive and scraping protocols for forming EBs generate EBs of wildly varying size, whereas the AggreWell methods generates are EBs with tightly controlled size distributions.

Because the number and size of the wells in each plate is the same large quantities of uniformly-size EBs can be formed, reducing variation between experiments.