STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit

Serum-free media for differentiation of human PSCs to ventricular cardiomyocytes and long-term maintenance of human PSC-derived cardiomyocytes

STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit

Serum-free media for differentiation of human PSCs to ventricular cardiomyocytes and long-term maintenance of human PSC-derived cardiomyocytes

STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit
1 Kit
700 USD
Catalog # 05010

Serum-free media for differentiation of human PSCs to cardiomyocytes and long-term maintenance of human PSC-derived cardiomyocytes

Product Advantages


  • Supports the entire hPSC-derived cardiomyocyte workflow

  • Simple monolayer protocol produces cardiomyocytes in 15 days

  • One serum-free kit generates over 50 million cardiomyocytes (cTnT+)

  • Robust performance with minimal variability across multiple hPSC lines

What's Included

  • STEMdiff™ Cardiomyocyte Differentiation Basal Medium, 380 mL
  • STEMdiff™ Ventricular Cardiomyocyte Differentiation Supplement A (10X), 10 mL
  • STEMdiff™ Ventricular Cardiomyocyte Differentiation Supplement B (10X), 10 mL
  • STEMdiff™ Ventricular Cardiomyocyte Differentiation Supplement C (10X), 20 mL
  • STEMdiff™ Cardiomyocyte Maintenance Basal Medium, 490 mL
  • STEMdiff™ Cardiomyocyte Maintenance Supplement (50X), 10 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

Overview

STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit (Catalog #05010) includes a medium for differentiation of human embryonic stem (ES) and induced pluripotent stem (iPS) cells (human pluripotent stem cells [hPSCs]) into ventricular cardiomyocytes (cardiac troponin T-positive [cTnT+]), as well as a medium for maintenance of hPSC-derived cardiomyocytes. This serum-free kit can be used to generate ventricular cardiomyocytes derived from a clump culture of hPSCs maintained in mTeSR™1 (Catalog #85850), mTeSR™ Plus (Catalog #100-0276), TeSR™-AOF (Catalog #100-0401), or TeSR™-E8™ (Catalog #05990). Greater than 80% of these cells will be cTnT+. An average of 1 x 10^6 cells can be harvested from a single well of a 12-well plate.

STEMdiff™ Cardiomyocyte Maintenance Kit (Catalog #05020) comprises the maintenance basal medium and supplement; it can be used for long-term maintenance of hPSC-derived cardiomyocytes for one month or longer. These cardiomyocytes can be used in various downstream applications and analyses.

NOTE: This product was formerly named ‘STEMdiff™ Cardiomyocyte Differentiation Kit’; the product itself and manufacturing procedures have not changed, but the name has been updated to more accurately reflect the cell type generated.
Subtype
Specialized Media
Cell Type
Cardiomyocytes, PSC-Derived
Species
Human
Application
Cell Culture, Differentiation, Maintenance
Brand
STEMdiff
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Stem Cell Biology
Formulation
Serum-Free

Data Figures

Figure 1. Cardiomyocyte Differentiation Protocol

Two days before the differentiation protocol, hPSC colonies are harvested and seeded as single cells at 350,000 cells/well in a 12-well format in TeSR™ medium. After one day (Day -1), the medium is replaced with fresh TeSR™ medium. The following day (Day 0), the TeSR™ medium is replaced with Medium A (STEMdiff™ Cardiomyocyte Differentiation Basal Medium containing Supplement A) to begin inducing the cells toward a cardiomyocyte fate. On day 2, a full medium change is performed with fresh Medium B (STEMdiff™ Cardiomyocyte Differentiation Basal Medium containing Supplement B). On days 4 and 6, full medium changes are performed with fresh Medium C (STEMdiff™ Cardiomyocyte Differentiation Basal Medium containing Supplement C). On day 8, medium is switched to STEMdiff™ Cardiomyocyte Maintenance Medium with full medium changes on days 10, 12 and 14, to promote further differentiation into cardiomyocyte cells. Small beating areas of cardiomyocytes can be seen as early as day 8, progressing to a full lawn of beating cardiomyocytes that can be harvested as early as day 15.

Figure 2. Morphology of hPSC-Derived Cardiomyocytes

Representative images of (A) hES (H9) cells and (B) hiPS (WLS-1C) cells on day 15 of differentiation to cardiomyocytes using the STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit. Differentiated cells exhibit typical cardiomyocyte morphology as an adherent, tightly packed web-like monolayer of beating cells. (C) Representative confocal microscopy image of a single hPSC-derived cardiomyocyte generated with the STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit and stained with cTnT (green) and DAPI (blue).

Figure 3. Efficient and Robust Generation of cTnT-Positive Cardiomyocytes

hES and hiPS cells were cultured for 15 days in single wells of 12-well plates using the STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit. At the end of the culture period, cells were harvested and analyzed by flow cytometry for expression of cardiac troponin T (cTnT). (A) Histogram analysis for cardiomyocyte cell marker cTnT for cultures of hES (H9) and hiPS (WLS-1C and STiPS-M001) cells. (Filled = sample; blank = secondary antibody only control) (B,C) Percentages and total numbers of cells expressing cTnT in cultures of hES or hiPS cells are shown. Data shown as mean ± SEM; n=3.

Figure 4. hPSC-Derived Cardiomyocytes Exhibit a Robust and Stable Excitability Profile

Microelectrode array (MEA) voltage recordings of cardiomyocytes (day 27) derived from human pluripotent stem cells generated and maintained with the STEMdiff™ Cardiomyocyte Differentiation and Maintenance Kits. The hPSC-derived cardiomyocytes have a characteristic electrical profile and stable beat rate. A large depolarization spike followed by a smaller repolarization deflection is observed.

Microelectrode array and flow cytometry of human ES and iPS cells maintained in mTeSR™1 (daily feeds) or mTeSR™ Plus (restricted feeds) and differentiated to cardiomyocytes using the STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit.

Figure 5. Generation of Cardiomyocytes from hPSCs Maintained in mTeSR™ Plus

Human ES (H9) and iPS (WLS-1C) cells were maintained in mTeSR™1 (daily feeds) or mTeSR™ Plus (restricted feeds) and differentiated to cardiomyocytes using the STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit. At the end of the differentiation period, cells were harvested and analyzed by microelectrode array (MEA) and flow cytometry. (A) Representative MEA voltage recordings of cardiomyocytes (day 20) demonstrate a characteristic electrical profile and stable beat rate. (B) Percentages of cells expressing cTNT and (C) total number of viable cells harvested are shown. Data are expressed as the mean (± SEM); n=2.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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Safety Data Sheet 1
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Safety Data Sheet 2
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Safety Data Sheet 6
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Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Resources and Publications

Publications (12)

Role of Blood Oxygen Saturation During Post-Natal Human Cardiomyocyte Cell Cycle Activities. L. Ye et al. JACC. Basic to translational science 2020 may

Abstract

Blood oxygen saturation (SaO2) is one of the most important environmental factors in clinical heart protection. This study used human heart samples and human induced pluripotent stem cell-cardiomyocytes (iPSC-CMs) to assess how SaO2 affects human CM cell cycle activities. The results showed that there were significantly more cell cycle markers in the moderate hypoxia group (SaO2: 75{\%} to 85{\%}) than in the other 2 groups (SaO2 {\textless}75{\%} or {\textgreater}85{\%}). In iPSC-CMs 15{\%} and 10{\%} oxygen (O2) treatment increased cell cycle markers, whereas 5{\%} and rapid change of O2 decreased the markers. Moderate hypoxia is beneficial to the cell cycle activities of post-natal human CMs.
Extracellular Vesicles from Skeletal Muscle Cells Efficiently Promote Myogenesis in Induced Pluripotent Stem Cells. D. Baci et al. Cells 2020 jun

Abstract

The recent advances, offered by cell therapy in the regenerative medicine field, offer a revolutionary potential for the development of innovative cures to restore compromised physiological functions or organs. Adult myogenic precursors, such as myoblasts or satellite cells, possess a marked regenerative capacity, but the exploitation of this potential still encounters significant challenges in clinical application, due to low rate of proliferation in vitro, as well as a reduced self-renewal capacity. In this scenario, induced pluripotent stem cells (iPSCs) can offer not only an inexhaustible source of cells for regenerative therapeutic approaches, but also a valuable alternative for in vitro modeling of patient-specific diseases. In this study we established a reliable protocol to induce the myogenic differentiation of iPSCs, generated from pericytes and fibroblasts, exploiting skeletal muscle-derived extracellular vesicles (EVs), in combination with chemically defined factors. This genetic integration-free approach generates functional skeletal myotubes maintaining the engraftment ability in vivo. Our results demonstrate evidence that EVs can act as biological shuttles" to deliver specific bioactive molecules for a successful transgene-free differentiation offering new opportunities for disease modeling and regenerative approaches."
Modeling Type 1 Diabetes In Vitro Using Human Pluripotent Stem Cells. N. C. Leite et al. Cell reports 2020 jul

Abstract

Understanding the root causes of autoimmune diseases is hampered by the inability to access relevant human tissues and identify the time of disease onset. To examine the interaction of immune cells and their cellular targets in type 1 diabetes, we differentiated human induced pluripotent stem cells into pancreatic endocrine cells, including $\beta$ cells. Here, we describe an in vitro platform that models features of human type 1 diabetes using stress-induced patient-derived endocrine cells and autologous immune cells. We demonstrate a cell-type-specific response by autologous immune cells against induced pluripotent stem cell-derived $\beta$ cells, along with a reduced effect on $\alpha$ cells. This approach represents a path to developing disease models that use patient-derived cells to predict the outcome of an autoimmune response.

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