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BMEM Protein-Free Cardiomyocyte Differentiation Kit

BMEM Protein-Free Cardiomyocyte Differentiation Kit

Optimized cardiomyocyte differentiation for human iPSC

Catalog #1003

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$110.00 USD
$110.00 USD
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BMEM Protein-Free Cardiomyocyte Differentiation Kit is a ready-to-use highly optimized, chemically defined, animal component-free kit for the differentiation of human induced pluripotent stem cells (hiPSC) to contracting cardiomyocytes (hiPSC-CM) in adherent culture.

This kit is a novel formulation optimized without any proteins. Differentiation using this kit typically produces contracting cardiomyocytes in 8-9 days and results in ~85-90% TNNT2+ cells and yields of >1x105 cells per cm2. Cardiomyocytes can be maintained in BMEM Cardiomyocyte Maintenance Medium for >45 days.

BMEM Protein-Free Cardiomyocyte Differentiation Kit is compatible with hiPSC growth medium BMEM Human Primed Pluripotent(1001).

BMEM Protein-Free Cardiomyocyte Differentiation Kit is compatible with a variety of culture matrices including: Corning® Matrigel® GFR (354230), Gibco™ Geltrex™ GFR (A1413202), R&D Systems™ Cultrex™ UltiMatrix GFR (BME001-05), Gibco™ Recombinant Vitronectin (A14700), and Corning® Synthemax II-SC (3535).

Each lot of BMEM Protein-Free Cardiomyocyte Differentiation Kit is performance tested on hiPSC and sterility tested.

Larger unit sizes are available on request

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Product Information

The following components are sold as part of the BMEM Protein-Free Cardiomyocyte Differentiation Kit and are not available separately.
Component Name Size Storage
BMEM Cardiomyocyte d0 100 mL Store at 4°C
BMEM Cardiomyocyte d1 100 mL Store at 4°C
BMEM Cardiomyocyte d2 100 mL Store at 4°C
BMEM Cardiomyocyte Maintenance 2 x 500 mL Store at 4°C

Instructions/Directions for Use

Instructions for Use

BMEM Protein-Free Cardiomyocyte Differentiation Kit is ready to use. Bottles can be used straight from the refrigerator (4 °C) and should not be warmed to RT or 37 °C prior to use. 

Directions for Use

hiPSC should be growing on a consistent schedule achieving 70-80% confluence before differentiation. Fast, consistent growth, without overgrowth (>90% confluence) is essential for reproducible differentiation. We recommend that hiPSC lines are ≥p20 for reproducible differentiation. For consistency, media changes should be completed at the same time of day.

Contracting cardiomyocytes should be seen after 8-9 days.

We recommend replating hiPSC-CMs to new wells for assays between d16 and d20.

Product Use

For Research Use Only. 

Not for diagnostic procedures. 

 Troubleshooting

High levels of cell death on d0-d1. 

Cause: hiPSC were over (>90%) or under (<50%) confluent prior to starting differentiation.

Solution: Adjust split ratio of pluripotent cells to achieve 70-80% confluency. Keep culture conditions consistent.

 Full contracting monolayer is not achieved.

Cause: hiPSC were under (<60%) or over (>90%) confluent prior to starting differentiation.

Solution: Adjust split ratio of pluripotent cells to achieve 70-80% confluency. Keep culture conditions consistent.

 No contracting cells by d10.

Cause: media change schedule deviated too far from prescribed protocol, potentially with inadequate exposure time to d0 media.

Solution: Make sure that media changes accurately follow the described schedule: 24 h d0, 24 h d1, 48 h d2.

Cause: hiPSC are growing poorly.

Solution: Grow cells until a reproducible split ratio every 4 days, as described above, is achieved. 

Cause: Media is too old.

Solution: Media should be used within 30 days of opening. 

Contracting monolayer constantly detaches from the plate surface. 

Cause: extracellular matrix is not suitable for long-term maintenance of hiPSC-CMs without passage, such as vitronectin or Synthemax.

Solution: Use an alternative matrix. If matrices such as vitronectin are desired to be used, then hiPSC-CMs should be passaged at d8-d10 when they can be replated into vitronectin.