CDI has made iPS cells from a variety of sources including fibroblasts and blood cells. The iPS cells used in the iCell Cardiomyocytes were derived by CDI from a commercially available, non-transformed, fibroblast cell line.
2. Can I provide the starting cell material to CDI for creation of a specific iPS cell line?
An iPS cell reprogramming service is currently under development. Contact CDI directly for further information.
3. What other cell types is CDI working on?
Our primary focus will be on additional pure sub-populations of the cardia lineage as well as hepatocytes, neural cells and cells of the hematopoietic lineage. CDI has several programs centered on additional cell types including hepatocytes,endothelial cells, neuronal cells, and blood cells.
4. From what organisms does CDI create stem cell-derived products?
Human stem cells are the focus of CDI’s operations, as they provide the most physiologically relevant model system for drug discovery and safety/toxicity assessment of human pharmaceuticals.
5. Can I purchase iPS cell lines directly from CDI?
Contact CDI directly for information on licensing iPS cell lines.
6. Do you perform routine karyotype analysis on your iPS starting material?
Yes, iPS cell cultures used for iCell Cardiomyocyte production are karyotyped by G-banding every 5 passages. All cells used for production have a normal female karyotype (46,XX).
1. What is CDI’s iCell Cardiomyocyte production and product availability?
CDI has industrialized the process of generating and differentiating stem cells into specific cell types. Our proprietary manufacturing process reliably produces sufficient quantities for large-scale pharmaceutical investigations.
2. How does CDI manufacture the iCell Cardiomyocytes?
CDI has a robust production process that is outlined in the figure below. This process yields pure cardiomyocytes in quantities that meet and exceed current cardiomyocyte demands.
3. How does CDI achieve such high purity for the iCell Cardiomyocytes?
Blasticidin is used to purify iCell Cardiomyoctyes from other differentiated cells. This process is highly efficient, with purity often exceeding 95% per cardiomyocyte lot.
4. How are iCell Cardiomyocytes shipped?
CDI ships iCell Cardiomyocytes on dry ice in cryovials as frozen suspensions of dissociated cells.
1. How do I best thaw iCell Cardiomyocytes?
The proper thawing and plating procedures are detailed in the iCell Cardiomyocytes User’s Guide.
2. Will one vial be enough to plate cells for a 6-well plate?
Plating density depends upon the projected use. A beating syncytial layer is recommended for most cell-based assays and requires approximately 47,000 plated cardiomyocytes per cm2. The following table lists the recommended numbers of plated cardiomyocytes to achieve a syncytial monolayer in commonly used flat-bottomed, multi-well plates.
| Plate Format (# of Wells) |
Well Area (cm2) |
Plated Cardiomyocytes/Well |
Plated Cardiomyocytes/Plate |
| 384 | 0.06 | 2,820 | 1,082,000 |
| 96 | 0.32 | 15,000 | 1,440,000 |
| 24 | 1.9 | 89,000 | 2,136,000 |
| 12 | 3.8 | 179,000 | 2,148,000 |
| 6 | 9.6 | 451,000 | 2,706,000 |
3. Can I use commercially available pre-coated plates for plating iCell Cardiomyocytes?
iCell Cardiomyocytes have successfully adhered to all commercially available pre-coated plating substrates tested to date, including: collagen I, collagen IV, and fibronectin. The substrate commonly used by CDI is 0.1% gelatin.
4. How soon should I expect to see contractile activity after thawing and plating iCell Cardiomyocytes?
Distinct areas will start beating 24 hours after plating while at 96 hours post-plating the cells will have formed a syncytium and will beat in synchrony if plated at a high enough density.
5. For my assay, I need to plate the iCell Cardiomyocytes at a high density. Can I concentrate the cells?
The iCell Cardiomyocytes User’s Guide details thawing and plating at the recommended density as well as higher and lower densities.
1. How often will I have to change the medium?
As outlined in the iCell Cardiomyocytes User’s Guide, the iCell Cardiomyocytes Maintenance Medium should be changed every other day.
2. Do I need to use antibiotics in the media to maintain the purity of my iCell Cardiomyocytes culture?
No. iCell Cardiomyocytes are supplied as a highly pure population of cardiomyocytes. Cell purity is maintained through the use of supplied media: iCell Cardiomyocyte Plating Medium and iCell Cardiomyocyte Maintenance Medium.
3. How often will I have to pass the iCell Cardiomyocytes?
iCell Cardiomyocytes do not require passaging.
4. If I plate the cells on a Friday, can I wait until Monday to change the medium from iCell Cardiomyocytes Plating Medium to iCell Cardiomyocytes Maintenance Medium?
The purpose of iCell Cardiomyocyte Plating Medium is to promote viability and attachment at thaw. To maintain iCell Cardiomyocytes health, the iCell Cardiomyocyte Plating Medium should be replaced with iCell Cardiomyocyte Maintenance Medium at 48 hours post-thaw. The proper feeding and maintenance procedure is described in the iCell Cardiomyocytes User’s Guide and the iCell Cardiomyocytes Training Video on CDI’s webpage.
5. How long do iCell Cardiomyocytes survive in culture, once initiated?
High-purity iCell Cardiomyocytes can survive for at least two weeks when cultured in iCell Cardiomyocytes Maintenance Medium, allowing you to perform chronic exposure assay and culture studies.
6. Can I maintain the iCell Cardiomyocytes at 5% CO2 instead of 7% CO2?
The bicarbonate level contained in the iCell Cardiomyocytes Maintenance Medium should provide proper pH conditions for culturing cells at CO2 levels ranging from 5% to 10%. Investigations are underway to confirm that iCell Cardiomyocytes perform optimally at 5% CO2.
1. Is special media required to culture iCell Cardiomyocytes?
CDI strongly recommends using iCell Cardiomyocytes Plating Medium and iCell Cardiomyocyte Maintenance Medium for culturing iCell Cardiomyocytes. These media have been specially formulated for optimal cell performance.
2. Do the iCell Cardiomyocytes media contain antibiotics?
Neither the iCell Cardiomyocyte Plating nor the Maintenance Media contain antibiotics. Therefore, sterile technique is crucial when handling iCell Cardiomyocytes.
3. Can I add antibiotics to the iCell Cardiomyocytes media?
Yes, antibiotics can be added to the iCell Cardiomyocytes Maintenance Medium as required by your experiments. We recommend using Gentamicin at a concentration of 25ug/mL.
4. Do the iCell Cardiomyocytes Maintenance or Plating Media contain any serum?
Yes, both media contain serum.
1. What cell source did CDI use to produce the iPS cell line from which iCell Cardiomyocytes are derived?
The iPS cell clone used to make the iCell Cardiomyocytes is a proprietary clone generated at CDI. This iPS cell clone was derived from cells which are available from a commercial source.
2. How many different genetic backgrounds are available with iCell Cardiomyocytes?
Currently, iCell Cardiomyocytes are available from one genetic background. Our product development teams are working to produce diverse genetic panels of cells for our customers.
3. What percentage atrial/ventricular/nodal phenotype cells are in iCell Cardiomyocytes?
CDI’s differentiation and selection process yields iCell Cardiomyocyte batches comprised of approximately 50% ventricular-like cells, with atrial- and nodal-like cells making up the remainder.
4. Can I purchase atrial, nodal and ventricular populations separately?
At this time, purified separate populations of atrial, ventricular, and nodal cells are not available. Our product development teams are working on methods to purify specific populations of cardiac cells from cardiomyocytes.
5. What are CDI’s criteria of characterization for iCell Cardiomyocytes?
CDI has extensive and specific criteria including purity, plating efficiency, electrophysiological activity, and responsiveness to known cardiotoxic compounds.
6. What ion channels are active in iCell Cardiomyocytes?
Automated and conventional patch clamp techniques have measured INa, Ito, If, IK1, IKr and ICa from iCell Cardiomyocytes.
7. Do the iCell Cardiomyocytes de-differentiate over time in culture?
No. Whole genome transcript analysis and functional phenotypic analysis suggests that the iCell Cardiomyocytes stably express the appropriate genes and behavior for at least 90 days past their release date.
8. Do iCell Cardiomyocytes beat spontaneously?
Yes. This spontaneous beating is an indication that the cells are healthy (see our video of beating iCell Cardiomyocytes).
9. Are iCell Cardiomyocytes tested for sterility, and specifically mycoplasma?
Yes. Each batch undergoes testing for bacterial and mycoplasma contamination.
10. What is the purity of iCell Cardiomyocytes, and how is it determined?
iCell Cardiomyocytes’ purity levels are routinely above 95%, determined by the percentage of cells in a batch that express red fluorescent protein under control of a cardiac-specific promoter.
11. What is the DMSO tolerance of iCell Cardiomyocytes?
iCell Cardiomyocytes are tolerant to up to 1% DMSO for 1 week.
1. Within what applications/assays have iCell Cardiomyocytes been tested?
iCell Cardiomyocytes have been tested in a variety of electrophysiological and cell-based applications/assays. Electrophysiological assays include conventional voltage and current patch clamp, automated voltage and current patch clamp, microelectrode array (MEA) analysis, and intracellular (sharp electrode) recordings. Cell-based assays include:
2. Can iCell Cardiomyocytes be transfected?
iCell Cardiomyocytes have been transfected using the Amaxa and Fugene HD systems. Transfection efficiencies of 20 - 30% have been achieved with the Amaxa protocol, while those achieved with Fugene HD were lower and may require protocol optimization.
3. iCell Cardiomyocytes contain RFP. Does this mRFP expression interfere with cell-based assays?
No. Our experience has been that the RFP levels are low enough to not have a significant impact when iCell Cardiomyocytes are used in cell-based assays.
