Stem cell expansion
Pluripotent stem cell expansion using either embryonic stem cells (ES) or induced pluripotent stem cells (iPSC) is a critical technique for producing cells used in disease modeling, drug discovery, therapeutic development. The expansion process involves the proliferation of ES/iPSCs to obtain sufficient quantities for research or translation to the clinic while maintaining the ES/iPSC’s pluripotency and differentiation capacity.
Problem
The culture of pluripotent stem cells, wether in an R&D tissue culture lab or a GMP cell manufacturing suite, is technically challenging requiring highly trained scientists.This person must exercise precise motor control when exchanging media or reagents and harvesting the cells, as well as make judgment decisions on when to initiate the above process steps. This is compounded by stem cells themselves being in a dynamically unstable cell state and can frequently suffer from loss of pluripotency, spontaneous differentiation, or cell death.
In plain English. You usually need someone with a PhD to move liquids between plastic containers who won’t mess up a long and complex number of cell culture steps.
This is undesirable due to:
- Inconsistency - Human variability, from the angle at which a pipette gun is held to the velocity of ejection of liquid from the stripette to deciding “what confluence” looks like creates points for poor process repeatability and failure.
- Contamination - poor aseptic technique, or honest mistakes, can contaminate cultures setting you back weeks.
- The need for Sleep - human beings generally don’t perform at their best when feeding cells at 3am.
- Boredom - Smart human beings thrive on solving interesting problems. Not being a machine. We predict having your team perform repetitive cell culture workflows will have negative effects on staff retention.
- Costs - have you ever tried to grow and manage a cell culture team? Operational expenses, equipment, and salaries shoot up fast. Surprisingly fast.
Solution
Your technical team creates value when planning/analyzing experiments. Not when they are feeding cells on the weekends. Liberate your team. Have a machine do the cell culture instead. Use a Unicorn cell manufacturing system.
Our cell culture automation and manufacturing systems pull together fluidics, mechanical actuation, in-line metabolic sensing and machine intelligence (not to jump on the AI bandwagon) to provide end-to-end automation of ES/iPSC expansion. From seeding and media and reagent exchange to passaging and harvesting.
Plug-and-play cell culture cartridges support conducting multiparameter experimental testing to determine ideal culture conditions, culturing up to 18 cell lines or testing 18 culture conditions in parallell, or using all cartridges to bulk up a single cell line.
Critically, our systems deliver unparalleled level of precision and standardization. From machine control of the tilt angle of cell culture vessels within 0.1 degrees to maintaining homogenous temperature distribution in the incubation chamber, and onboard metabolite analytics. Reduce variability. Mitigate contamination concerns. And reclaim your peace of mind knowing that our systems will culture your cells while you sleep. What more could you ask for?
Proof
In a benchmarking study, Emmet reliably expanded iPSCs that maintained their pluripotency.
An anchorage dependent iPSC expansion protocol was selected, where 1 million iPSCs would be expanded to 500 million iPSCs over a 10 day expansion period. The initial 1 million iPSCs were simultaneously seeded into a cell culture cartridge within Emmet (machine) and a control tissue culture flask that was cultured manually (manual).
During initial seeding and subsequent passages, Emmet demonstrated increased homogeneity of seeding when compared to manual cell control, as evidenced by this heat map of representative cell confluence measured before initiating a passage.
Whether your protocol calls for twice hourly, or twice weekly, media exchanges, Emmet can automate your reagent transfers, feeding cycles as well as the process steps to dissociate, passage and harvest cells.
On day 10 of the expansion protocol. After multiple passages and reaching the final desired cell count, a final dissociation was initiated and expanded iPSCs were harvested from Emmet and the manual control.
The number of cells produced by Emmet was consistent with the control throughout, and iPSCs expanded both in Emmet and in the manual control had similar expression levels of pluripotency markers, indicating that both expanded iPSC populations retained their pluripotency.
