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2010 - Cor.At Cardiomyocytes: Primary-like Cardiomyocytes for Manual and Automated Electrophysiological Screening

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Lonza Resource Notes (2010)

Authors:
Kettenhofen R., Stölzle S.

Source: 
Lonza Resource Notes (2010) Fall:9-10


Abstract: 

Cor.At® Cardiomyocytes are derived from mouse embryonic stem cells (mESC). During differentiation of the mESC, about 5% of all cells develop into cardiomyocytes. Using transgenic mESC with the puromycin resistance cassette under the control of the cardiac α-myosin heavy chain-(MHC) promoter, 99.9% pure Cor.At® Cardiomyocytes can be selected from the large amount of noncardiac myocyte cell population by the application of puromycin. For long-term storage, Cor.At® Cells are deep frozen as single cell suspensions in liquid nitrogen or -150°C deep freezers. Quality control strategies are implemented to guarantee lot-to-lot reproducibility and uniformity of functional properties of Cor.At® Cardiomyocytes for a storage period of at least 12 months. Thawed Cor.At®  Cardiomyocytes readily form spontaneously and synchronously contracting monolayers overnight.
Seeded in low density on cover slips, Cor.At® Cardiomyocytes can be applied to manual patch clamp for the recording of action potentials as well as all three typical cardiac ion currents INa, ICa,L and IK (data not shown). Additionally, single cell suspensions of pre-cultured  Cor.At® Cardiomyocytes can be readily analyzed with very high success rates in automated patch clamp systems like the Port-a-Patch® and Patchliner® from Nanion Technologies GmbH, Munich, Germany, as well as in other automated patch clamp systems (data not shown). The uniqueness of both Nanion systems is their capability to record action potentials in the current clamp mode and the possibility to perform the recordings at physiological temperature in addition to the standard measurements of ion currents in the voltage clamp mode.


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