ReproCardio2

Product Classification : Human iPS Cells Derived CardiomyocytesResearch Theme : Cardiac disease, Drug Effect Test, Pharmacological Effectiveness Test, Electrophysiology Testing, Safety Test, Toxicology test

ReproCardio
summary

Human iPS Cell Derived Cardiomyocytes

Features

 ・For efficient in vitro cardiotoxicity assays
 ・Functionally characterized using various reference compounds
 ・Expresses cardiac ion channels and cardiomyocyte-specific markers


Overview

Convenient frozen cells for wide range of applications

ReproCardio2 cardiomyocytes are delivered as frozen single cells and can be re-constituted into spheroids in embryoid body formation inducing 3D culture technologies like low adherence plates. Reconstituted ReproCardio2 clumps beat periodically and robustly which provide excellent in vitro models of in vivo heart.

Characterization

ReproCardio2™ cells express various cardiomyocyte-specific markers and cardiac ion channels (see images below).

MLC-2A

MLC-2A

Myosin light chain specific for the atrium of the mammalian heart.

MLC-2V

MLC-2V

Myosin light chain specific for the ventricle of the mammalian heart.

alpha-MHC

alpha-MHC

Myosin heavy chain which is mainly expressed in the atrium of the heart.

beta-MHC

beta-MHC

Myosin heavy chain beta which is mainly expressed in the ventricle of the heart.

cTnT

cTnT

Cardiac troponin T, a component of troponin complex necessary for contraction

Cx43

Cx43

Connexin 43, the main cardiac connexin found mainly in ventricular heart cells.

These photos were taken with Yokogawa live cell imaging system“CellVoyager CV1000″.
CellVoyager is a registered trademark of Yokogawa Electric Corporation.

Product details

Catalog No. Product Price
RCDC001N ReproCardio2 Contact us

Contents & storage conditions

RCDC001N ReproCardio2 contents

  • human iPS cell-derived single cardiomyocytes
    1 x 106 cells
Storage conditions ReproCardio2 cells: liquid nitrogen
ReproCardio Culture Medium 2: -20 ℃
After thawing the medium, keep at 4 ℃ and use within 2 weeks.
ReproCoat (coating solution): Room temperature or 4 ℃

Related Products

Catalog No. Product Price
RCDC101 ReproCardio2 Culture Medium 2 (100 ml) Contact us
RCDC301 ReproCardio2 Assay Medium (100 ml) Contact us
RCHEOT004 ReproCoat Contact us
RCESD005 QTempo assay service Contact us

Electrophysiological characterization with patch clamp

ReproCardio2 can be used for manual patch clamp experiments when cultured as single cells. in vivo-like activities and properties including spontaneous action potential, Na/Ca/K channel currents are reliably observed.
(Click on the figures to enlarge)

Spontaneous action potential (current clamp recording)

Spontaneous action potential (current clamp recording)

Sodium current (voltage clamp recording)

Sodium current (voltage clamp recording)

Calcium current (voltage clamp recording)

Calcium current (voltage clamp recording)

Potassium current (voltage clamp recording)

Potassium current (voltage clamp recording)

Data and images courtesy of DSTC (Drug Safety Testing Center Co, Ltd.)
株式会社薬物安全性試験センター

Use ReproCardio 2 as thin layers

*ReproCardio Assay Medium is highly recommended for assays.
ReproCELL has developed new protocols for thin layer culture of ReproCardio 2.
MEA extracellular recording and optical calcium imaging protocol are available upon request.

MEA recordings from beating thin layer formed by ReproCardio 2

When plated on a flat surface coated with ReproCELL's coating material provided in the kit, ReproCardio 2 cardiomyocytes form a thin layer and start beating. Extracellular electrical recordings can be made by using MEA, and clear sodium and potassium peaks can be observed.

MEA recording from a thin layer constituted of ReproCardio 2 cardiomyocytes. Right panel: Extracellular electrical activity. Red vertical lines indicates sodium and potassium signals.

※We recommend that the CO2 concentration (5%) is fixed during a measurement with MEA for acquiring stable data.

Incubator box for fixing the CO2 concentration

Incubator box for fixing the CO2 concentration

Live imaging of calcium concentration change using ReproCardio 2 thin layer

Calcium wave propagation over the cardiomyocyte thin layer can be observed by using conventional calcium-sensitive dyes. The accessibility of the cells within a thin layer for drugs makes this suitable for testing the effects of compounds to calcium activity of cardiomyocytes.

Calcium imaging of a thin layer constituted of ReproCardio 2 cardiomyocytes using a calcium-sensitive dye (fluo-8, ABD Bioquest). Right panel: quantification of fluorescent intensity which represents calcium dynamics in the cells.

Use ReproCardio 2 as clumps

*ReproCardio Assay Medium is highly recommended for assays.

※ Data is provided by Cyfuse Biomedical.


Bio 3D printer (Regenova®) and
Cardiac-like structure fabricated using ReproCardio2


cyfuse

ReproCardio2 (ReproCELL, Inc.) are cells derived from human iPSC. When cultured, these cells give rise to highly mature, beating, cardiomyoctyes. By combining these cells with others and using the Regenova Bio-3D-Printer (Cyfuse Biomedical K.K.), human cardiac-like structures can be created. Advanced 3D structures like these, fabricated with the Regenova instrument, provide new approaches and applications for innovative in vitro assays and regenerative medicine.




≪Construction of cardiac-like structure│How to fabricate≫



≪Construction of cardiac-like structure│Observation of movement≫



≪Construction of a cardiac-like structure│immunostaining and tissue staining≫

The cells in the cardiac-like structure show specific localization that depends in the cell types.

Protocol

Instruction manual for RCDC001N (Jump to the download link, open in a new window)

Presentations

  • Society for Biomolecular Sciences(SBS), 14th AnnualConference & Exhibition 2008 (St. Louis), poster session, April 2008.
  • IBC Asia's 4th Annual Stem Cells Asia Congress (Singapore), oral session, June 2008.
  • ELRIG and SBS Present: Drug Discovery (UK) poster session, September 2008.
  • Advances in Stem Cell Discoveries (San Francisco) oral session, September 2008.
  • Stem Cells: Drug Discovery and Therapeutics (London) oral session, February 2008.
  • Society for Biomolecular Science 15th Annual Conference (France), poster session, Best Poster 2009 Award-winning, April 2009.
  • Stem Cells and Regenerative Medicine Europe (UK), September 2009, Best Poster 2009 Award-winning.
  • MipTec Conference 2009, October 13-15, Basel Invited Lecture.
  • The 74th Annual Scientific Meeting of the Japanese Circulation Society (Kyoto), March 2010.
  • The 130th The Pharmaceutical Society of Japan (Okayama), March 2010.
  • The Society of Toxicology (SOT) 51st Annual Meeting (CA, USA), poster session, March 2012.
  • Safety Pharmacology Society (SPS), 13th annual meeting 2013 (Rotterdam, Netherlands), poster session, September 2013.
  • Automated Patch-Clamp systems and Ion Channel Expressing Cells 2013 (Tokyo), Verbal presentation 2013 Nov.
  • The 7th Takeda Science Foundation Symposium (Osaka), Poster presentation 2014 Jan.

Publications

ReproCardio, ReproCardio2

  • Datta-Chaudhuri, Timir, Pamela Abshire, and Elisabeth Smela. "Packaging commercial CMOS chips for lab on a chip integration." Lab on a chip 14.10 (2014): 1753-1766.
  • Datta-Chaudhuri, Timir, Pamela Abshire, and Elisabeth Smela. "Packaging commercial CMOS chips for lab on a chip integration." Lab on a chip 14.10 (2014): 1753-1766.
  • Mandenius, Carl-Fredrik, and Thomas Meyer. "High-throughput screening assays to evaluate the cardiotoxic potential of drugs." High-Throughput Screening Methods in Toxicity Testing (2013): 403-420.
  • Scott, Clay W., Matthew F. Peters, and Yvonne P. Dragan. "Human induced pluripotent stem cells and their use in drug discovery for toxicity testing." Toxicology letters 219.1 (2013): 49-58.
  • Takeuchi, Akimasa, et al. "Microfabricated device for co-culture of sympathetic neuron and iPS-derived cardiomyocytes." Engineering in Medicine and Biology Society (EMBC), 2013 35th Annual International Conference of the IEEE. IEEE, 2013.
  • Asai, Yasuyuki, et al. "Combination of functional cardiomyocytes derived from human stem cells and a highly-efficient microelectrode array system: an ideal hybrid model assay for drug development." Current stem cell research & therapy 5.3 (2010): 227-232.
  • Asai, Y. "Direct measurement of the QT interval in stem cell-derived cardiomyocytes for the assessment of QT liability." Nihon yakurigaku zasshi. Folia pharmacologica Japonica 134.6 (2009): 320-324.
  • Szebenyi, Kornelia, et al. "Human pluripotent stem cells in pharmacological and toxicological screening: new perspectives for personalized medicine." Personalized Medicine 8.3 (2011): 347-364.

QTempo assay

  • Vidarsson, Hilmar, Johan Hyllner, and Peter Sartipy. "Differentiation of human embryonic stem cells to cardiomyocytes for in vitro and in vivo applications."Stem Cell Reviews and Reports 6.1 (2010): 108-120.
  • Lecina, Marti, et al. "Scalable platform for human embryonic stem cell differentiation to cardiomyocytes in suspended microcarrier cultures." Tissue Engineering Part C: Methods 16.6 (2010): 1609-1619.
  • Phillips, Blaine W., and Jeremy M. Crook. "Pluripotent Human Stem Cells."BioDrugs 24.2 (2010): 99-108.