Differentiation
Differentiating any type of cell, be it iPSC or mesenchymal stromal cells, is a crucial workflow for producing the desired end cell type you need for use in basic research or therapeutic. Differentiation protocols generally involve sequentially applying growth factors or small molecules at precise time or event driven points to your cell culture.
Problem
Differentiating cells is technically challenging often requiring highly trained scientists to execute complicated workflows while ensuring precise fluid velocity and homogenous fluid distribution across multiple cell culture vessels to prevent aberrant differentiation.
These manual workflows are undesirable due to:
- Inconsistency - Within a top-notch group of five TC scientists, there will be five different ways to distribute differentiation media across a flask, and five different estimations of when the 'right' time to perform media exchanges is.
- Environmental Variability - Every condition, from whether or not you have air conditioning in your lab, to the effects of thermally cycling your reagents can affect the performance of your differentiation protocol.
- Complex Tech Transfers - No amount of SOP writing will ever capture the precise conditions under which a protocol was first developed. This can cause months to years of delays when transferring protocols to different facilities or cell culture teams.
- Upset Team Members- in our experience, people don’t like working on a differentiation protocol that calls for frequent media exchanges for weeks unless they’re purposefully trying to burn themselves out.
- Costs - Have you ever ordered replacement growth factors to re-do a failed differentiation protocol? Or have you calculated how much staff time is spent conducting differentiations? Expenses compound at an alarmingly fast rate.
In plain English. Humans acting as the production systems to differentiate cells is bonkers - this has to change.
Solution
Your technical team creates value when planning and analyzing experiments. Not when they are performing endless amounts of manual differentiations. Have Emmet do your differentiations instead.
Emmet leverages machine-driven fluid control, mechanical actuation and onboard reagent storage, in-line sensing, and machine intelligence-driven process control. This mitigates the effects of reagent thermal cycling, removes human variability during media exchanges, and provides quality control beyond "what color is the media" and "do the cells look right."
Emmet brings an unmatched level of precision and standardization to your cell culture while you remain in the loop, able to pause machine actions at any time. Are you ready for your TC to join the 21st-century
The Proof
In a benchmarking study, Emmet reliably differentiated iPSCs down a mesoderm lineage towards adipocytes.
An adipogenic differentiation protocol was selected where iPSCs were clump passaged into embryoid bodies (EBs), expanded for two days in suspension conditions, and then passaged into plasma-treated cell culture vessels for cell outgrowth and the initiation of adipogenic differentiation. The end-to-end protocol was conducted in Emmet.
During the initial stage of the protocol, Emmet was able to form EB’s without any operator assistance. Subsequent passaging to non-plasma treated (for EB growth in suspension culture conditions) and back to plasma treated (for re-plating of cells and outgrowth in adherent conditions) culture vessels were again, performed entirely by Emmet. Subsequently, application of growth factors and a cocktail of small molecules was applied to initiate adipogenic differentiation.
This process could be run in reverse. To clump passage Emmet expanded iPSCs into EBs, and transfer the EBs to a suspension culture vessel for additional proliferation and differentiation in suspension conditions.
On day 14 after the expression of the adipogenic phenotype was observed, a dissociation was initiated and the differentiated adipocytes were harvested from Emmet and the manual control.
The cells differentiated by Emmet contained intracellular lipid droplets, a typical phenotype of adipocytes and had high expression levels of late stage adipogenic differentiation markers.
Finally, we compared triglyceride expression levels of the iPSC derived adipocytes Emmet generated with the triglyceride expression levels of two othe cell types we had Emmet produce in a separate experiment: immortalised pre-adipocyte derived stem cell (imADSCs) and machine differentiated imADSCs. All three Emmet cultivated cell types showed expected triglyceride expression levels.
All experiments performed by Unicorn Biotechnologies scientists at Unicorn's Sheffield, UK R&D lab on an Emmet: Generation 1 instrument.
