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iCell CardioSpheres² Kit, 01434
Products
Kits
- 0.5M iCell Cardiac Fibroblasts, 01434
- 1.25M iCell Cardiomyocytes², 01434
- 0.5M iCell Endothelial Cells, 01434
- 30ml iCell Cardiomyocytes Plating Medium
- 100ml iCell Cardiomyocytes Maintenance Medium
- 25ml iCell Cardiac Co-Culture Supplement A
- 100μl iCell Plating Supplement B (1000X)
- 250μl iCell Cardiac Co-Culture Supplement B
genetic status
Quantity (Cells Per Vials)
Catalog #
Catalog #:
CardioSpheres² differentiated from human iPS cells, frozen
From
$3,186.00
Product Overview
Three-dimensional (3D) in vitro human heart models offer a biologically relevant platform for studying cardiac function and disease. While cardiomyocytes make up only about 50% of the cells in the left ventricle, incorporating additional cell types, such as endothelial cells and cardiac fibroblasts, into multicellular systems enhances physiological relevance, predictive power, and mechanistic insight. Using human iPSC technology, these cell types can be derived from the same donor and cryopreserved for co-culture. When assembled into 3D microtissues, these co-cultures provide a straightforward and effective approach to mimic native heart tissue, serving as a complementary tool to traditional cardiomyocyte-only models.
iCell® CardioSpheres2 are human 3D cardiac microtissues engineered by co-culturing human iPSC-derived iCell Cardiomyocytes2, iCell Endothelial Cells, and iCell Cardiac Fibroblasts in an optimized co-culture medium right out of thaw without the need for pre-plating or gel matrix. Using FCDI’s Application Protocol, these cells self-assemble in ultra-low attachment multi-well plates to form tri-culture cardiac spheroids. This in vitro model mimics key features of native heart tissue in vivo, enabling iCell CardioSpheres2 to support testing of structural integrity (maturity, organization, cell alignment) and functional output (contraction, electrical activity, drug response) using high content imagers, thus offering a reliable tool for studying cardiac physiology and pharmacology in a high throughput format.
Components:
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Need Help With This Product?
Our specialists are here to help you find the best product for your application.
Our regular business hours are 9:00am to 5:00pm Central Time (USA)
Technical Docs
PROTOCOLS
Application Protocols
Generation of 3D iCell CardioSpheres and iCell CardioSpheres² with iCell Products
USER DOCS
Quick Guide
iCell Cardiomyocytes2, 01434 Quick Guide
Datasheet
iCell CardioSpheres2 Datasheet
Biosafety Document
iCell Cardiomyocytes and iCell Cardiomyocytes2 01434 Biosafety Document
Safety Data Sheet
iCell Plating Supplement B Safety Data Sheet
Safety Data Sheet
iCell Cardiac Co-Culture Supplement B Safety Data Sheet – Asia
Safety Data Sheet
iCell Cardiac Co-Culture Supplement B Safety Data Sheet
Safety Data Sheet
iCell Cardiac Co-Culture Supplement A Safety Data Sheet – Asia
Safety Data Sheet
iCell Cardiac Co-Culture Supplement A Safety Data Sheet
Safety Data Sheet
iCell Cardiomyocytes Maintenance Medium, iCell Cardiomyocytes Plating Medium Safety Data Sheet - Germany
Performance Data
Protocol for Generation of iCell CardioSpheres 01434
Workflow schematic for generating 3D cardiac spheroids. Human iPSC-derived cell types, including iCell Cardiomyocytes, iCell Cardiac Fibroblasts, and iCell Endothelial Cells from donors 01434 or 11713 were supplied by FUJIFILM Cellular Dynamics, Inc. (FCDI). iCell Cardiomyocytes Maintenance Medium (iCMM), as well as the 5X iCell Cardiac Co-Culture Supplement, were also from FCDI. All three cell types were thawed and combined at a ratio of 65:15:20 into ultra-low attachment (ULA) plates (e.g. PrimeSurface® 96w or 384w from Sbio). 3D microtissues were maintained for ~14 days in culture until ready for testing. On the day of assay, cells were loaded with EarlyTox™ Cardiotoxicity calcium dye (Molecular Devices) for 2 h and then imaged on a high-throughput cellular screening system, such as the FLIPR Penta or FDSS/mCELL. Spontaneous calcium oscillations were recorded using 30-50 frames per second (fps) that allowed for resolution of complex cardiac waveforms.
Figure 1: Workflow to generate and assay iCell CardioSpheres
Figure 2: iCell Cardiomyocytes2 batches maintain consistently high purity.
Characterization of iCell Cardiomyocytes2
The iCell Cardiomyocytes2 are a highly pure population, all lots are >95% positive for cardiac marker, cardiac troponin T. This is confirmed by immunostaining, and flow cytometry at thaw (Day 0) and after 8 days in culture. The cardiac troponin T expression across multiple manufacturing batches with high purity demonstrate the consistently and reproducible manufacturability of the iCell Cardiomyocytes2.
Characterization of iCell Endothelial Cells
iCell Endothelial Cells have the appropriate cobble stone morphology and are >90% pure. They endothelial cells express markers such as CD31 (PECAM), CD105 (Endoglin) and CD144 (VE-Cadherin) with appropriate pattern of localization for Zo-1, Von Willebrand Factor (vWF) and CD144 (VE-Cadherin).
Figure 3: iCell Endothelial Cells are >90% pure and express key endothelial markers with appropriate pattern of localization.
Figure 4: iCell Cardiac Fibroblasts have high purity and express key fibroblast specific markers.
Characterization of iCell Cardiac Fibroblasts
iCell Cardiac Fibroblasts have the appropriate flat and spindle shaped morphology and are >90% pure. These iCell Cardiac Fibroblasts express markers such as TE7 and are quiescent (negative for αSMA), with appropriate pattern of localization for COL1A1, Cx43 and POSTN.
iCell CardioSpheres: High Throughput Screening Compatible
Combining iCell Cardiomyocytes, iCell endothelial cells and iCell Cardiac Fibroblasts directly from thaw into ultra-low attachment plates following the application protocol generates uniform single CarioSpheres in 96-well and 384-well format using 10,000 or 5,000 total cells respectively. This makes iCell CardioSpheres amenable to high throughput screening assays.
Figure 5:
Figure 6: iCell CardioSpheres are spontaneous beating and display uniform calcium oscillations when loaded with calcium dye (EarlyTox or Calcium 6) making them amenable for high throughput screening assays.
Baseline Cardiac Activity in Calcium Oscillation Assay
iCell CardioSpheres generated using our application protocol generates one spheroid per well of 96w or 384w plate. These CardioSpheres also demonstrate spontaneous beating and display uniform calcium oscillations across different wells when loaded with calcium (fluorescent green when bound to calcium) dye (EarlyTox or Calcium 6) making them amenable for high throughput screening assays in 96 or 384 well format.
Cardioactive Compound Pharmacology on CardioSpheres
iCell CardioSpheres respond to compounds known to modulate cardiac activity, including E-4031 (hERG channel blocker), Bay K 8644 (calcium channel activator), Isoproterenol and Digoxin (positive inotrope), and Verapamil (calcium channel blocker) when compared to vehicle control (DMSO 0.1%)
Figure 7: iCell CardioSpheres respond to compounds known to modulate cardiac activity, including E-4031 (hERG channel blocker), Bay K 8644 (calcium channel activator), Isoproterenol and Digoxin (positive inotrope), and Verapamil (calcium channel blocker).
Product Highlights
iCell CardioSpheres2 Highlights
- Easy to implement – iCell Cardiomyocytes2, endothelial cells and fibroblasts are shipped cryopreserved with optimized media. Simply thaw and use.
- Human cells – iCell CardioSpheres2 cell components are from the same donor hence isogenic.
- Highly pure – iCell products that are part of the CardioSpheres2 Kit are highly pure and enable assembly of CardioSpheres2 in a matrix/substrate free manner in ultra-low attachment (ULA) plates.
- Homogenous and reproducible – Generate single iCell CardioSpheres2 using the 3 cell types to generate homogenous and physiologically relevant microtissue that can provide biologically relevant and reproducible results.
- High throughput Screening – iCell CardioSpheres2 can be assembled in 96 or 384 well ULA plates and can be used for high throughput screening functional assays.