Perfused Organ Panel Starter System, with 48-Well Plates
Perfused Organ Panel Starter System
PerfusionPalTM Organ-on-a-Chip insert system Product Content Template
PerfusionPal provides physiologically relevant interstitial perfusion and superior oxygenation to 3D cell cultures cultured in the wells of a 12-well or a 48-well insert. This translates to enhanced cell viability and function for pharmaceutical, CRO, biotech and academic customers. PerfusionPal’s superiority has been proven in weeks-long maintenance of complex primary models of brain and liver, for example, with remarkable outcomes resulting in two-fold to five-fold increase in cellular respiratory metabolism and log-fold increase in Phase I drug metabolism relative to 2D and 3D controls.
PerfusionPal enables seamless integration into routine workflow with superior outcomes in both short-term and long-term studies. It requires attachment of just one tube to perfuse 48 statistically independent organ models, all in their own media. Drugs and assay reagents are added and removed as in any multiwell plate. Assays may be run in situ, supernatants transferred to another plate for plate reader readouts, and cultures removed and transferred to another dish for imaging.
Perfused Organ Panel is a unique system that enables continuous perfusion of 48 statistically independent organ cultures with just one tube. Media and assays are added and removed as in a standard 48-well plate. Scaffolds can be removed for cell isolation and imaging.
Starter system includes:
- A dual syringe pump
- Two 48-well PerfusionPal plates
- Two syringes and tubing
- 120 SeedEZ scaffolds
- Two bottles of 100 mL blood substitute
With the exception of syringe pump, all components are sterile. The blood substitute is immiscible with and insoluble in media, reagents, and drugs. Its main functions are to enable perfusion and provide superior gas exchange to the basal side of each culture. The blood substitute does not support the growth of microorganisms. It is the only chemical in the syringes and the tubing. The risk of contamination is therefore negligible compared to other perfusion platforms that utilize the culture media in tubing and syringes.
Features and Benefits include:
- Parallel perfusion of 96 cultures (48 cultures in each plate)
- Superior gas exchange
- In-well perfusion (concentrated secretome and metabolome remains in the well)
- Routine media changes (using micropipette as in any multiwell plate)
- In-well assaying
- Assay multiplexing (e.g. bioluminescence and fluorescence)
- Routine reads (transfer supernatants to a new plate and read on a plate reader)
- Extended mechanistic studies
- Repeat drug dose testing
- Increased Phase I drug metabolism
- Human metabolites for slow-to-metabolize drugs
- Concentrated secretome for testing biologics (monoclonal antibodies and antibody-drug conjugates)
- Saves time, and money spent on cells in other assays (such as relay assay)
What solution does it provide to the customer?
Extended cell survival and extended functional outcomes from days to weeks even for primary cells
This enables researchers to answer questions that cannot be asked today due to limited capabilities of other in vitro culturing methods, for example:
- poor survival of primary cells in vitro due to inadequate oxygenation
- contact inhibition in 2D cell cultures that limits duration of studies to 4-5 days, or
- a necrotic core and ischemia in a 3D spheroid model that leads to culture decay
Superior culturing conditions, provided by PerfusionPal, translate to reproducible results for scientific research, cancer research, drug testing and translational research, enabling, for example, the testing of personalized drug strategies using patients’ cells.
Multi-fold increase in drug metabolism for ADMET research
Without human drug metabolites, the drug toxicity may not be known until early human trials.
Approximately 30% of new drugs metabolize slowly and their metabolites cannot be obtained using human cells in vitro. Animal testing is not very predictive. Animals have different drug metabolizing enzymes and therefore, metabolites. Specifically, the CYP450 isoforms for 5 out of 7 the most important drug metabolizing enzymes are different between a human and animal species.
Culturing cells in PerfusionPal increases the activity of drug metabolizing enzymes. This increase in drug metabolism on a per cell basis is further enhanced by an extended ability of cells to metabolize drugs when cultured in PerfusionPal. Both conditions generate an opportunity to obtain human drug metabolites in vitro and to detect them easily from a concentrated metabolome provided by in-well perfusion. Once human metabolites are obtained and identified as non-reactive or toxic, the data may be fed into standard physiological software such as Simcyp to predict the in vivo clearance from the in vitro data; thus, saving time and money, while reducing risks to volunteers in clinical trials.
Vascularized tumors for cancer research
The majority of cancer drugs are used to treat patients which tumors are vascularized and metastasized, yet the majority of tumor models for cancer research represent avascular tumors, an early stage in disease development before the tumor develops its own blood supply and continues to grow.
PerfusionPal enables long-term growth of advanced tumor models with heterogeneous cell populations, extracellular matrix, and intra-tumor supply of nutrients for disease-state relevant small-molecule drug and biopharmaceutical testing.
Testing of high molecular weight biopharmaceuticals in vitro
Biopharmaceuticals, such as humanized monoclonal antibodies and antibody-drug-conjugates, are active only in humans. They are frontline therapies for cancer, immune, inflammatory, infectious and neurological diseases. These drugs are injected intravenously, distributed by vasculature and clear slowly, from days to weeks. Current in vitro methods cannot support cultures long enough to provide physiologically relevant clearance times, or tissue-like resistance to interstitial delivery and distribution of high MW therapeutics for their adequate testing.
PerfusionPal supports high-cell-density advanced tissue models and enables interstitial delivery and distribution of high MW biopharmaceuticals for their adequate long-term testing in vitro, using human cells in culture. Furthermore, in-well perfusion provides concentrated secretome, providing researchers with an opportunity to detect cytokine release, responsible for severe reactions in clinical trials.
A simple-to-use organ-on-a-chip compatible with standard laboratory equipment
Continuously perfused, organ-on-a-chip systems comprise 1-2 organs with no replicates and require expensive microscopes and incubation systems for continuous monitoring of culture activity.
PerfusionPal supports 48 organs with one tube. It enables researchers to parallelize studies, multiplex assays as in any 48-well plate, and allows use of low-cost laboratory equipment, such as plate reader, to read results in a minute. It provides an alternative to 48 studies with 48 serial chips, and 48 expensive microscopes with 48 people required to monitor culture activity. Most importantly, PerfusionPal is reliable; there are no microchannels to clog, and cultures can be maintained long-term for repeat drug dose testing or mechanistic research.
What does the product do better than other products/solutions on the market? Identify the key USP
Commercially available cell culture perfusion products and products in development satisfy only one of the two necessary requirements of commercial customers, either function or throughput:
- Continuous perfusion devices have throughput = 1 or are low throughput.
- High-throughput devices that do not provide perfusion.
PerfusionPal is the only product capable of perfusing 48 statistically independent organs in their own media with 1 tube and 1 pump. No other system can do this without 96 tubes, 48 pumps, and 48 media bottles for 48 organs, and certainly not without room-sized incubators to support all the peripherals these devices require.
Realistically, CN Bio Innovations’ and Mimetas’ products are in multiwell format, and therefore the most advanced relative to single chips. However, a built in pump in each well of CN Bio Innovations product complicates its use. There are too many wires/ cables to connect and controllers to supervise. PerfusionPal’s blood substitute and one tube solve all these problems.
Mimetas product has a higher throughput than our PerfusionPal, but our PerfusionPal brings superior gas exchange to cultures. This is important, because cells just like ourselves dye faster from the lack of gas than the lack of nutrients.
What are the other benefits of using the product?
- PerfusionPal is simple to setup and use. It requires attachment of one tube to perfuse 48 cultures.
- The use of blood substitute as a pumping liquid, eliminates the need for 48 pumps, 48 media bottles and 96 tubes that would otherwise be used in a 48-well organ-on-a-chip system.
- In PerfusionPal, media, drugs and assay reagents are added and removed using micropipette as in any multiwall plate.
- In enclosed organs-on-a-chip, drugs are typically injected and removed through special ports with separate tubes or microfluidics; this is not trivial to do.
- PerfusionPal has reliable and consistent performance.
- In contrast with other organ-on-a-chip technologies, in PerfusionPal there is no clogging of microchannels, no failures in operation, no need to service the system while running, and no drug adsorption to the interior of small channels contributing to uncertainty in drug concentration.
- PerfusionPal is the only perfusion system with zero dead volume in fluidics.
- In PerfusionPal, media and drugs are not wasted in pumps, tubes, media bottles etc. as in all other perfusion technologies. Instead, media and drugs are added to the well and stay in the well during perfusion. This saves money in expensive developmental drugs and biologics.
- PerfusionPal provides superior oxygenation and therefore, cellular function.
- This is an open system with two gas exchange interfaces; air at the top and blood substitute at the bottom of the culture. All other organ-on-a-chip and perfusion systems are generally enclosed and oxygen availability is low.
- Biomarker discovery/ diagnostic and prognostic functions
- In-well perfusion provides concentrated secretome, metabolome, and signalling molecules that may be concentrated enough to facilitate detection of new biomarkers of therapeutic efficacy or therapy surveiilance.
- Small molecule drug testing (ADME/Tox)
- Biopharmaceutical testing (on target efficacy/ off-target toxicity of antibody-drug-conjugates and multi-specific antibodies)
- Cancer research
- Academic research
Data and examples of use
Phase I drug metabolism Log-fold increased baseline CYP Activity in Primary Human Hepatocytes after 7 days in culture
Baseline metabolism was recorded for various CYP enzymes in primary human hepatocytes (BioIVT Cryoplateable 10-donor Liverpool) in cells cultured in 2D or PerfusionPal. Asterisk indicate significant difference in metabolic activity due to plating condition. Cells cultured in the Lena Biosciences PerfusionPal system show an enhanced CYP activity compared to the control. All values were normalized to the 2D control. Note: Log Scale for baseline activity graph. *p<0.05, **p<0.01, ***p<0.001.
Increased CYP activity in Induced Primary Human Hepatocytes cultured with PerfusionPal
Phase 1 Drug Metabolism in primary human hepatocytes (BioIVT Cryoplateable 10-donor Liverpool) was measured over a 7 day period. Cells were cultured in either a 2D format using a multi-well plate, or in the Lena Biosciences PerfusionPal system (n=3 for all conditions). At day 6, cells were induced with either Rifampin (10 uM) or Omeprazole (50 mM) and data was collected on day 7.
Perfusion rate for the PerfusionPal data corresponds to 16 volume changes per 24 hour period.
All data was normalized by their respective un-induced data (shown in black and green). Asterisk indicate significant enhancement of induced hepatocytes in the PerfusionPal system when compared to the control. All induced data was normalized to baseline CYP activity *p<0.05, **p<0.01, ***p<0.001.
Culturing human hepatocytes in the PerfusionPal consistently enhances the baseline activity of the key drug metabolizing enzyme, CYP1A.
Prior to EROD assays, HepaRG™ cells, iPSC-derived Hepatocytes 2.0 (Cellular Dynamics) and a 10-donor pool primary hepatocytes (BioreclamationIVT Cryoplateable Liverpool)™ were maintained in culture for 5 days. HepG2 cells, that had significantly lower activity from all other cell types, were maintained for 7 days. The results were normalized to respective 2D controls for each cell type. 2D refers to 2D cell cultures, 3D refers to 3D cultures cultured in SeedEZ, 3D static refers to 3D cultures cultured in SeedEZ in unperfused PerfusionPal with the blood substitute (the effect of superior oxygenation), and 3D perfused refers to 3D cultures cultured in SeedEZ and perfused in PerfusionPal. *p<0.05, **p<0.01, ***p<0.001
Increase in cellular respiratory metabolism in PerfusionPal for HepG2 liver cell line, HepaRG differentiated hepatobiliary cells, and BT-474 HER2+ breast cancer cell line.
Long-term culture of complex culture models
Primary brain tri-culture model and differentiated liver hepatobiliary model perfused in parallel in PerfusionPal for 4 weeks prior to imaging; and breast cancer model at 10X
How it works:
- The cells seeded within the SeedEZ scaffold are perfused by their culture medium. They receive oxygen supply and gas exchange both apically and basally, by employing Blood Substitute as a liquid piston.
- Although the bottom of each well is open to the blood substitute, the individual wells in our PerfusionPal are prohibited from mixing with one another. Our careful and patented design allows for different cells and different media conditions to be utilized in different wells by restricting each of these conditions to the well.
- Once the scaffolds are seeded with cells and medium added, a perfusion rate is set, and PerfusionPal plate is placed in the incubator. Media can be changed and tested for metabolites or cytokines while the study is taking place.
- The risk of contamination is negligible. Compared to other perfusion setups that use culture media in tubes, the only liquid in the PerfusionPal tube is the Blood Substitute that, unlike culture media, does not support the growth of microorganisms.
- Once studies are complete, culture media is removed with a pipette and discarded. The Blood Substitute that remains in the tray is collected, sterile filtered, and reused for subsequent studies.
|Description||PerfusionPal starter system with 48 well plates|