Gibco™ PSC Cardiomyocyte Differentiation Kit

Catalog No.	Quantity
A2921201	1 kit

Catalog No. A2921201 Supplier Gibco™ Supplier No. A2921201

Description

PSC Cardiomyocyte Differentiation Kit

The Gibco™ PSC Cardiomyocyte Differentiation Kit consists of a set of serum-free and xeno-free media that enable efficient differentiation of human pluripotent stem cells (PSCs) to contracting cardiomyocytes in as few as 8 days. Unlike other methods that require multiple components and longer assay duration, the PSC Cardiomyocyte Differentiation Kit can generate cardiomyocytes from pluripotent stem cells in a ready-to-use media format and in less time.

Comprised of three 1X media that require no thawing or mixing, each medium is used consecutively over a total of 14 days, resulting in functional cardiomyocytes that express relevant physiological markers, contract in culture, and can be subsequently maintained in culture for >15 days (maintenance medium also sold separately (Cat. No. A2920801)).

No thawing or mixing required
The PSC Cardiomyocyte Differentiation Kit is comprised of 1X media that do not require frozen storage, so no thawing or mixing of reagents. Simply warm the media and add to PSC culture.

Rapid production of cardiomyocytes
The PSC Cardiomyocyte Differentiation Kit is capable of producing contracting cardiomyocytes in as few as 8 days. Differentiated cardiomyocytes can be subsequently maintained in culture for >15 days.

Generation of high quality cardiomyocytes
Cardiomyocytes generated using the PSC Cardiomyocyte Differentiation Kit are functionally relevant, expressing key markers such as TNNT2, Nkx2.5, MYH6, and α-actinin; and contract in culture.

Specifications

• Cell Type: Cardiomyocytes
• Content And Storage: • 500 mL Cardiomyocyte Maintenance Medium
  • 100 mL Cardiomyocyte Differentiation Medium A
  • 100 mL Cardiomyocyte Differentiation Medium B
  
  Store at 2–8° and protect from light.
• Product Type: PSC Cardiomyocyte Differentiation Kit
• Quantity: 1 kit
• Shipping Condition: Room Temperature

Frequently Asked Questions (FAQs)

What substrate should I use for cardiomyocyte differentiation?

We recommend using Gibco Geltrex Flex LDEV-Free hESC-qualified Reduced Growth Factor Basement Membrane Matrix (Cat. No. A4000046801, A4000046802, or A4000046803) or Gibco Vitronectin (VTN-N) Recombinant Human Protein, Truncated (Cat. No. A400457 or A400458) for xeno-free applications.

Is StemFlex Medium compatible with downstream differentiation kits available from Thermo Fisher Scientific?

Yes. We have seen compatibility with the following differentiation kits provided by Thermo Fisher Scientific: PSC Cardiomyocyte Differentiation Kit (Cat. No. A2921201), PSC Definitive Endoderm Induction Kit (Cat. No. A3062601), PSC Neural Induction Medium (Cat. No. A1647801), and PSC Dopaminergic Neuron Differentiation Kit (Cat. No. A3147701).

I used the Gibco PSC Cardiomyocyte Differentiation Kit and am not seeing good differentiation with my iPSC line. What should I do?

We recommend always using H9 or H7 ESC line as a control in your experiments. We recommend adjusting the cell density or extending the induction time for difficult-to-differentiate iPSC lines.

I am planning to use the PSC Cardiomyocyte Differentiation Kit. Which markers can I expected my differentiated cells to express?

Cardiomyocytes generated using PSC Cardiomyocyte Differentiation Kit have been tested for key markers such as TNNT2, Nkx2.5, MYH6, and Alpha-Actinin. The Cardiomyocyte Immunocytochemistry Kit (Cat. No. A25973) contains validated antibodies to measure TNNT2 and Nkx2.5 in cultures generated using PSC Cardiomyocyte Differentiation Kit.

How long can I maintain differentiated cells in culture using the Gibco PSC Cardiomyocyte Differentiation Kit?

Differentiated cells can be maintained for a month or longer for long-term studies. We recommend the use of Gibco Geltrex Matrix for long-term cultures.

Which population of cardiomyocytes is produced using the PSC Cardiomyocyte Differentiation Kit?

The population of cardiomyocytes produced is a mix of atrial and ventricular cells. Over time, cultures become more ventricular.

What dissociation reagent do you recommend using for passaging during differentiation of PSCs to cardiomyocytes?

We recommend using EDTA for passaging PSCs and Gibco TrypLE Enzyme for dissociating cardiomyocytes prior to cryopreservation.

How many plates of cardiomyocytes can be generated from one PSC Cardiomyocyte Differentiation Kit?

Each kit contains enough volume for eight 12-well plates for 14 days of culture.

What substrate should I use for cardiomyocyte differentiation?

We recommend using Gibco Geltrex LDEV-Free, hESC Qualified Reduced Growth Factor Basement Membrane Matrix (Cat. No. A1413301 or A1413302) or Gibco Vitronectin (VTN-N) Recombinant Human Protein, Truncated (Cat. No. A14700) for xeno-free applications.

Can the media supplied in the Gibco PSC Cardiomyocyte Differentiation Kit be frozen?

We do not recommend freezing the media.

Is it normal to see line to line variability in cardiac differentiation?

Yes. Variability is normal and it is not uncommon to find certain lines that will not differentiate as efficiently. Including a control line, such as the human ESC H1 or H9 cells, which have been shown to differentiate consistently well, may be helpful.

Can I purchase additional Cardiomyocyte Maintenance Medium without buying the entire Gibco PSC Cardiomyocyte Differentiation Kit?

Yes. Cardiomyocyte Maintenance Medium is sold separately as Cat. No. A22920801.

How long can I maintain differentiated cells in culture using the Gibco; PSC Cardiomyocyte Differentiation Kit?

Differentiated cells can be maintained for a month or longer for long-term studies. We recommend the use of Gibco Geltrex Matrix for long term cultures.

I am planning to use the Gibco PSC Cardiomyocyte Differentiation Kit. What markers can I expected my differentiated cells to express?

Cardiomyocytes generated using PSC Cardiomyocyte Differentiation Kit have been tested for key markers such as TNNT2, Nkx2.5, MYH6, and ?-Actinin. The Cardiomyocyte Immunocytochemistry Kit contains validated antibodies to measure TNNT2 and Nkx2.5 in cultures generated using PSC Cardiomyocyte Differentiation Kit.

What density of PSCs should I start the cardiomyocyte differentiation protocol with?

Our studies have shown that the confluency of cells when starting differentiation can have a dramatic impact on efficiency of differentiation. Therefore, we recommend a range finding study to determine optimal confluency of each PSC line when starting differentiation. Guidance for this can be found in the product insert. Additionally, we recommend singularizing cells prior to differentiation rather than passaging in small clumps. Singularizing PSCs for differentiation to cardiomyocytes allows better seeding and confluence estimates, resulting in more consistent results well-to-well and overall better differentiation of difficult to differentiate lines.

What are the critical parameters in my starting PSCs that influence the cardiomyocyte differentiation outcome?

It is critical to use high quality human PSCs (with minimal or no differentiated colonies) that are karyotypically normal, confirmed to exhibit pluripotency markers, and are undergoing routine culture with regular subculture intervals and maintaining healthy morphology before starting cardiomyocyte differentiation. Additionally, we recommend that PSC line not be used past 100 passages.

How many cardiomyocytes can I produce from my starting population of PSCs?

In general, yields will be 100-200 fold the starting population of PSCs. For example, from 0.6 million PSCs per 12-well plate, you can produce 65-70 million cardiomyocytes.

Is there an optimal plate format for the Gibco PSC Cardiomyocyte Differentiation Kit?

The ideal plate format is 12-well due to the enhanced ability to create beating syncytium, easy harvesting, and characterization.

What is the purpose of each individual medium in the Gibco PSC Cardiomyocyte Differentiation Kit?

Cardiomyocyte Differentiation Medium A: Pushes PSCs toward mesodermal commitment via BMP/activin pathway activation and glycogen kinase 3 inhibition

Cardiomyocyte Differentiation Medium B: Induces cardiac mesoderm via Wnt inhibition

Cardiomyocyte Maintenance Medium: Matures cardiomyocytes

For Research Use Only. Not for use in diagnostic procedures.