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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. Please 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, or below
CDI has several programs centered on additional cell types including hepatocytes, 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?
Please 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 iCell Cardiomyocytes lot.
4. 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
5. 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.
6. How are iCell Cardiomyocytes shipped?
CDI ships iCell Cardiomyocytes on dry ice in cryovials as frozen suspensions of dissociated cells.
7. What percentage atrial/ventricular/nodal phenotype cells are in iCell Cardiomyocytes?
CDI’s differentiation and selection process yields iCell Cardiomyocytes batches comprised of approximately 50% ventricular-like cells, with atrial- and nodal-like cells making up the remainder.
8. Can I purchase atrial, nodal and ventricular populations separately?
At this time, purified separate populations of atrial, ventricular, and nodal cells are not available. CDI has programs working on methods to purify specific populations of cardiac cells from cardiomyocytes.
9. What are CDI’s criteria of iCell Cardiomyocyte characterization?
CDI has extensive and specific criteria including purity, plating efficiency, electrophysiological activity, and responsiveness to known cardiotoxic compounds.
10. 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.
11. Do iCell Cardiomyocytes proliferate?
iCell Cardiomyocytes are terminally differentiated and therefore do not proliferate. Proliferation of the small percentage of non-cardiomyocyte cells is prevented through the use of CDI’s proprietary culture media.
12. 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.
13. What is the DMSO tolerance of iCell Cardiomyocytes?
iCell Cardiomyocytes are tolerant to up to 1% DMSO for 1 week.
14. Do I need any specialized equipment to maintain iCell Cardiomyocytes?
Standard cell culture equipment is sufficient for maintaining iCell Cardiomyocytes. However, it is recommended that incubator CO2 levels be set to 7%.
15. 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.
16. Do iCell Cardiomyocytes beat spontaneously?
Yes. This spontaneous beating is an indication that the cells are healthy (see movie).
17. Are iCell Cardiomyocytes tested for sterility, and specifically mycoplasma?
Yes. Each batch undergoes testing for bacterial and mycoplasma contamination.
18. 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.
19. Within what applications/assays have iCell Cardiomyocytes been tested?
iCell Cardiomycoytes 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 Cell-Titer Glo Luminescent Cell Viability Assay, Caspase-Glo 3/7 Assay, CellTiter-Fluor Cell Viability Assay, MultiTox-Fluor Multiplex Cytotoxicity Assay (Promega), Cell Meter JC-10 Mitochondrial Membrane Potential Assay (ABD Bioquest), and CyQUANT Direct Cell Proliferation Assay (Invitrogen).
20. 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 Cardiomycoytes are used in cell-based assays.