hts of ca 2 transients in human ips derived
play

HTS of Ca 2+ Transients in Human iPS-derived Cardiomyocytes as a - PowerPoint PPT Presentation

Sep 29, 2023 •451 likes •871 views

Use our discoveries to advance yours HTS of Ca 2+ Transients in Human iPS-derived Cardiomyocytes as a Predictive and Cost Effective Assay Early on in Drug Development Dr. Ralf Kettenhofen Hamamatsu 10th European Functional

  1. Use ¡our ¡discoveries ¡to ¡advance ¡yours HTS of Ca 2+ Transients in Human iPS-derived Cardiomyocytes as a Predictive and Cost Effective Assay Early on in Drug Development Dr. Ralf Kettenhofen � Hamamatsu 10th European Functional Drug Screening Symposium � Axiogenesis AG, Cologne, Germany Actelion, Allschwil, Switzerland

  2. Content • Introduction into the production of Cor.4U cardiomyocytes • Overview of established applications with Cor.4U cardiomyocytes • HTS calcium transient assay in the Hamamatsu FDSS/ µ Cell • Compound screening • Data analysis • Outlook 2

  3. Generation of Cor.4U human iPS derived Cardiomyocytes • iPS generated according to Yamanaka (licensed from iPS Academia) • Production system by Axiogenesis enables large lot sizes • Tightly controlled differentiation results in minimal lot to lot variations • Selection based purification technology results in pure cardiomyocytes • Cryopreserved in various formats • GFP positive and colourless varieties will be available • Complete FTO for commercial use on the limited use label license • Cryopreserved • Fresh cultures on different substrates (cell culture flasks, 96/384 well plates) 3

  4. 4

  5. Use ¡our ¡discoveries ¡to ¡advance ¡yours Introduction Recording of Calcium Transients in Cor.4U Cardiomyocytes

  6. Excitation-Contraction Coupling Action Potential Currents Na + Amplitude Ca 2+ L-type Ca 2+ T-type Na + /Ca 2+ -exchanger K + I to K + IK s K + IK r Time K + IK ur Many Channel Types contribute to the Calcium is the messenger that integrates the Action Potential electrochemcial signals of the action potential with the molecular signaling pathways that regulate contraction

  7. Hamamatsu FDSS/ µ Cell

  8. Plating Efficiency on 384 Well Plates Fluo-4 Assay Data was kindly provided by Dr. Thomas Licher, Sanofi Germany

  9. FDSS Parameters Analysed by the CalDio/Wave Checker Software (3) P-P duration time (1)Peak Number (2)P rate(BPM) (6) A M P (5) RMP=Fluorescence Bottom intensity at the bottom peak (7) Max_slope[Fluorescence counts/ms] same as upstroke slope AMP= Peak Fluorescence count- RMP (but bottom to peak) (8) MaxNeg_slope[Fluorescence (4)RATIO = AMP/RMP counts/ms] same as downstroke slope (but peak to bottom) (9) APD 10 to 90 (1) Peak number (total, BPM) APD10 (2) P-P time [ms] (Ave, Std, Max, Min) APD50(Peak Width, FWHM) (3) Ratio (Ave, Std) � Ratio = (AMP+RMP)/RMP A M (4) AMP (Ave, Std) P (5) RMP (Ave, Std) (6) Slope (Ave, Std) � Rising Slope: Slope from bottom to peak � Falling Slope: Slope from peak to bottom � APD90 PWD 90 0% - 10%, 10% - 90%, 20% - 80%, 30% - 70% (7) Integration (Ave, Std) (8) - (16) PWD (10% - 90%) [ms] (Ave, Std)

  10. Use ¡our ¡discoveries ¡to ¡advance ¡yours hERG/I Kr blocker

  11. Use our discoveries to advance yours Astemizole hERG Blocker

  12. 5 min FDSS µ Cell - Astemizole vehicle control 41 nM 370 nM

  13. 30 min FDSS µ Cell - Astemizole vehicle control 4.57 nM 13.7 nM

  14. 30 min FDSS µ Cell - Astemizole 41 nM 123 nM 370 nM

  15. Use our discoveries to advance yours Data Analysis with the Wave Checker Module Astemizole

  16. FDSS µ Cell - Astemizole Wave Checker Module

  17. FDSS µ Cell - Astemizole

  18. FDSS µ Cell - Astemizole

  19. FDSS µ Cell - Astemizole export of text file format

  20. FDSS µ Cell - Astemizole

  21. FDSS µ Cell - Astemizole

  22. FDSS µ Cell - Astemizole

  23. FDSS µ Cell - Astemizole 45,00# 14,00# 40,00# average&falling&slope&[RFU/ms]& 12,00# bea$ng'frequency'[bpm]' 35,00# baseline# 10,00# baseline# 30,00# 5#min# 5#min# 8,00# 25,00# 10#min# 10#min# 20,00# 6,00# 15#min# 15#min# 15,00# 4,00# 20#min# 20#min# 10,00# 2,00# 25#min# 25#min# 5,00# 30#min# 30#min# 0,00# 0,00# 10000,00# 3333,33# 1111,11# 370,37# 123,46# 41,15# 13,72# 4,57# 1,52# 0,51# Control# 35#min# 10000# 3333# 1111# 370# 123# 41# 14# 5# 2# 1# Control# 35#min# concentra$on'[nM]' concentra7on&[nM]&

  24. FDSS µ Cell - Astemizole 18000,00# 14000,00# 16000,00# average&peak&width&60%&[ms]& 12000,00# 14000,00# average&peak&width&90%&[ms]& 10000,00# baseline# 12000,00# baseline# 5#min# 8000,00# 10000,00# 5#min# 10#min# 6000,00# 10#min# 8000,00# 15#min# 15#min# 4000,00# 6000,00# 20#min# 20#min# 4000,00# 2000,00# 25#min# 25#min# 30#min# 2000,00# 0,00# 30#min# 35#min# 10000,00# 3333,33# 1111,11# 370,37# 123,46# 41,15# 13,72# 4,57# 1,52# 0,51# Control# 0,00# 35#min# 10000,00# 3333,33# 1111,11# 370,37# 123,46# 41,15# 13,72# 4,57# 1,52# 0,51# Control# concentra8on&[nM]& concentra8on&[nM]& 14000,00# 14000,00# average&peak&width&80%&[ms]& 12000,00# 12000,00# average&peak&width&70%&[ms]& 10000,00# 10000,00# baseline# baseline# 5#min# 8000,00# 8000,00# 5#min# 10#min# 10#min# 6000,00# 6000,00# 15#min# 15#min# 4000,00# 4000,00# 20#min# 20#min# 2000,00# 25#min# 25#min# 2000,00# 30#min# 0,00# 30#min# # # # # # # # # # # # 35#min# 0 3 1 7 6 5 2 7 2 1 l o 0,00# 0 3 1 3 4 1 7 5 5 5 r , , , , , , , , , , t 35#min# 0 3 1 0 3 1 4 1 0 3 n 0 3 1 7 2 4 1 # # # # # # # # # # # o 0 3 1 7 6 5 2 7 2 1 l 0 3 1 3 1 o 0 3 1 3 4 1 7 5 5 5 C r 0 3 1 , , , , , , , , , , t 0 3 1 0 3 1 3 4 1 0 n 1 0 3 1 7 2 4 1 o 0 3 1 3 1 C 0 3 1 1 concentra8on&[nM]& concentra8on&[nM]&

  25. Use our discoveries to advance yours Dofetilide hERG Blocker

  26. 5 min FDSS µ Cell - Dofetilide vehicle control 24.7 nM 222 nM

  27. 30 min FDSS µ Cell - Dofetilide vehicle control 24.7 nM

  28. FDSS µ Cell - Dofetilide 70.00# 60.00# bea$ng'frequency'[bpm]' 50.00# baseline# 40.00# 5#min# 10#min# 30.00# 15#min# 20#min# 20.00# 25#min# 10.00# 30#min# 35#min# 0.00# # # # # # # # # # # # # l 3 0 0 1 4 3 9 7 2 7 0 o 0 1 3 9 7 2 6 0 2 6 0 r t . . . . . . . . . . . 0 0 0 0 2 8 4 4 2 6 n 0 2 7 2 6 0 o 2 6 0 C 2 concentra$on'[nM]' 10.00# 35.00# 9.00# average&falling&slope&[RFU/ms]& 30.00# 8.00# average&rising&slope&[RFU/ms]& 7.00# 25.00# baseline# 6.00# baseline# 20.00# 5#min# 5.00# 5#min# 10#min# 15.00# 10#min# 4.00# 15#min# 15#min# 3.00# 10.00# 20#min# 20#min# 2.00# 25#min# 5.00# 1.00# 25#min# 30#min# 0.00# 0.00# 30#min# Control# 0.03# 0.10# 0.30# 0.91# 2.74# 8.23# 24.69# 74.07# 222.22# 666.67# 2000.00# # # # # # # # # # # # # l 3 0 0 1 4 3 9 7 2 7 0 o 0 1 3 9 7 2 6 0 2 6 0 r . . . . . . . . . . . t 0 0 0 0 2 8 4 4 2 6 0 n 2 7 2 6 0 o 2 6 0 C 2 concentra6on&[nM]& concentra7on&[nM]&

  29. FDSS µ Cell - Dofetilide 800.00# 1600.00# 700.00# 1400.00# average&peak&width&30%&[ms]& average&peak&width&50%&[ms]& 600.00# 1200.00# baseline# baseline# 500.00# 1000.00# 5#min# 5#min# 400.00# 800.00# 10#min# 10#min# 300.00# 600.00# 15#min# 15#min# 200.00# 400.00# 20#min# 20#min# 100.00# 200.00# 25#min# 25#min# 0.00# 0.00# 30#min# 30#min# Control# 0.91# 24.69# 74.07# 222.22# 666.67# 2000.00# 0.03# 0.10# 0.30# 2.74# 8.23# Control# 0.03# 0.10# 0.30# 0.91# 2.74# 8.23# 24.69# 74.07# 222.22# 666.67# 2000.00# concentra8on&[nM]& concentra8on&[nM]& 14000.00# 5000.00# 4500.00# average&peak&width&70%&[ms]& 12000.00# average&peak&width&90%&[ms]& 4000.00# 10000.00# 3500.00# baseline# baseline# 3000.00# 8000.00# 5#min# 5#min# 2500.00# 10#min# 6000.00# 10#min# 2000.00# 15#min# 15#min# 1500.00# 4000.00# 1000.00# 20#min# 20#min# 2000.00# 500.00# 25#min# 25#min# 0.00# 0.00# 30#min# 30#min# Control# 0.03# 0.10# 0.30# 0.91# 2.74# 8.23# 24.69# 74.07# 222.22# 666.67# 2000.00# Control# 0.03# 0.10# 0.30# 0.91# 2.74# 8.23# 24.69# 74.07# 222.22# 666.67# 2000.00# concentra8on&[nM]& concentra8on&[nM]&

  30. FDSS µCell - Clustering of Compounds Confidential

  31. Outlook

  32. Outlook - Hamamatsu FDSS 7000EX Camera for fast recording of fluorescence and luminescence.

  33. Outlook Evaluation of different kinds of fluorescent calcium and membrane 
 potential dyes for the Cor.4U Cardiomyocytes Establishment of cardiac cytotoxicity/viability assay Combination of different luminescence assays viability cell death detection apoptosis 


  34. Dopaminergic and Sensory Neurons

  35. Patch Clamp of hiPS-derived Dopamineric Neurons Manual current clamp recording Manual voltage clamp recording of spontaneous action potentials of sodium currents 0.06 0.04 0.02 0.00 U[V] -0.02 -0.04 -0.06 -0.08 0 2 4 6 8 10 12 T[s] Data was kindly provided by PoreGenic GmbH, Rostock, Germany

  36. Patch Clamp of hiPS-derived Peripheral Neurons Manual voltage clamp recording of sodium currents raw data I/V diagram 1.00E-009 0.00E+000 -1.00E-009 I[A] -2.00E-009 -3.00E-009 -4.00E-009 -0.06 -0.04 -0.02 0.00 0.02 0.04 0.06 0.08 U[V] Data was kindly provided by PoreGenic GmbH, Rostock, Germany

Download Presentation
Download Policy: The content available on the website is offered to you 'AS IS' for your personal information and use only. It cannot be commercialized, licensed, or distributed on other websites without prior consent from the author. To download a presentation, simply click this link. If you encounter any difficulties during the download process, it's possible that the publisher has removed the file from their server.

Recommend

Functional Enhancement of Human iPSC-derived Cardiomyocytes Enabling Assessment of Inotropic

Functional Enhancement of Human iPSC-derived Cardiomyocytes Enabling Assessment of Inotropic

Functional Enhancement of Human iPSC-derived Cardiomyocytes Enabling Assessment of Inotropic Compounds and Improved Prediction of Compound Risk Yama Abassi, PhD Vice President of Global Strategic Business Development and Grow th ACEA

709 views • 21 slides
Understanding High Speed Surges/Transients TVA PQ Group - Transmission August 1, 2017

Understanding High Speed Surges/Transients TVA PQ Group - Transmission August 1, 2017

Understanding High Speed Surges/Transients TVA PQ Group - Transmission August 1, 2017 High-Speed Transients Source of Energy Discussed or Not Discussed Today Lightning Discussed Line or cable switching Discussed Nuclear Bomb Detonation

1.22k views • 79 slides
Human-centered Computing Lab Contextual Inference and Characterization Derived from Wireless Data

Human-centered Computing Lab Contextual Inference and Characterization Derived from Wireless Data

Human-centered Computing Lab Contextual Inference and Characterization Derived from Wireless Data Mining Rute Sofia (rute.sofia@ulusofona.pt) http://copelabs.ulusofona.pt Agenda Background and COPELABS R&D Unit 2017 Projects and

565 views • 22 slides
Astroinformatics in the Time Domain: Classification of Light Curves and Transients Prof. S.

Astroinformatics in the Time Domain: Classification of Light Curves and Transients Prof. S.

Astroinformatics in the Time Domain: Classification of Light Curves and Transients Prof. S. George Djorgovski With: M. Graham, A. Mahabal, A. Drake, and many students and collaborators Center for Data-Driven Discovery and Astronomy Dept.,

543 views • 50 slides
Analysis of the Effects of Single Event Transients on an SAR-ADC based on Charge Redistribution

Analysis of the Effects of Single Event Transients on an SAR-ADC based on Charge Redistribution

Analysis of the Effects of Single Event Transients on an SAR-ADC based on Charge Redistribution Graduate Program Department of Electrical Engineering Federal University of Rio Grande do Sul - UFRGS Alisson J. C. Lanot, Tiago R. Balen

445 views • 18 slides
Multimessenger Transients and the Liverpool Telescope A report on work during the first Advanced

Multimessenger Transients and the Liverpool Telescope A report on work during the first Advanced

Multimessenger Transients and the Liverpool Telescope A report on work during the first Advanced LIGO science run Chris Copperwheat Liverpool Telescope group: S.Bates, N.Clay, J.Marchant, C.Mottram, A.Piascik, R.Smith, I.Steele Collaborators:

448 views • 18 slides
Selecting Transients Automatically for the Identification of Models for an Oil Well IFAC Workshop

Selecting Transients Automatically for the Identification of Models for an Oil Well IFAC Workshop

Selecting Transients Automatically for the Identification of Models for an Oil Well IFAC Workshop on Automatic Control in Offshore Oil and Gas Production onio H. Ribeiro 1 Luis A. Aguirre 1 Ant Laboratory of Modeling, Analysis and Control of

275 views • 27 slides
Extreme Transients in the Multimessenger Era Philipp Msta Einstein fellow @ UC Berkeley

Extreme Transients in the Multimessenger Era Philipp Msta Einstein fellow @ UC Berkeley

Extreme Transients in the Multimessenger Era Philipp Msta Einstein fellow @ UC Berkeley pmoesta@berkeley.edu BlueWBlueWaters Symposium 2018 Sunriver Resort Core-collapse supernovae neutrinos turbulence (Binary) black holes accretion

812 views • 63 slides
IceCube as a Neutrino Follow-up Observatory for Astronomical Transients Kevin Meagher

IceCube as a Neutrino Follow-up Observatory for Astronomical Transients Kevin Meagher

IceCube as a Neutrino Follow-up Observatory for Astronomical Transients Kevin Meagher Universit libre de Bruxelles On Behalf of the IceCube Collaboration July 17, 2017 35 th International Cosmic Ray Conference Busan, Korea Overview

410 views • 21 slides
CSE 143 Substituting Derived Classes Recall that an instance of a derived class can always

CSE 143 Substituting Derived Classes Recall that an instance of a derived class can always

CSE 143 Substituting Derived Classes Recall that an instance of a derived class can always //client function (not a method) be substituted for an Dynamic Dispatch and Virtual void printPoint( Point pt ) instance of a base class {

196 views • 4 slides
there is no species of reasoning more common, more useful, and even necessary to human

there is no species of reasoning more common, more useful, and even necessary to human

there is no species of reasoning more common, more useful, and even necessary to human life, than that which is derived from the testimony of men. Evidentialism: A wise man proportions his belief to the evidence. A

454 views • 14 slides
High frequency transients produced during disconnector operations when switching small capacitive

High frequency transients produced during disconnector operations when switching small capacitive

High frequency transients produced during disconnector operations when switching small capacitive currents and impacts on instrument transformers Atlanta, Georgia, March 22, 2016 Presented by: Pierre Riffon Eng. Consultant, representing

483 views • 20 slides
Notes on derived categories and motives Daniel Krashen Table of Contents Introduction The

Notes on derived categories and motives Daniel Krashen Table of Contents Introduction The

Notes on derived categories and motives Daniel Krashen Table of Contents Introduction The bounded derived category of a variety Quick and dirty derived categories Triangulated categories Homotopy categories and derived categories Comparison

994 views • 71 slides
Workshop on Transients by C.M. Greenfield for Raffi Nazikian, Mark Foster, and a cast of many

Workshop on Transients by C.M. Greenfield for Raffi Nazikian, Mark Foster, and a cast of many

Workshop on Transients by C.M. Greenfield for Raffi Nazikian, Mark Foster, and a cast of many Presented at the Theory and Simulation of Disruptions Workshop July 15, 2015 DE-FC02-04ER54698 0

478 views • 23 slides
Derived Classes and Inheritance Chapter 9 D&D Derived Classes It is sometimes the case

Derived Classes and Inheritance Chapter 9 D&D Derived Classes It is sometimes the case

Derived Classes and Inheritance Chapter 9 D&D Derived Classes It is sometimes the case that we have a class is nearly what we need. Derived classes acquire the properties of an existing class. The original class is called the

512 views • 25 slides
Disk winds from NS merger remnants: EM transients & r-process nucleosynthesis Rodrigo

Disk winds from NS merger remnants: EM transients & r-process nucleosynthesis Rodrigo

6 4 2 0 -2 -4 -6 0 2 4 6 8 10 12 Disk winds from NS merger remnants: EM transients & r-process nucleosynthesis Rodrigo Fernndez UC Berkeley Dan Kasen, Eliot Quataert, & Josiah Schwab (UC Berkeley) Brian Metzger

305 views • 18 slides
The Quality Assurance Act for construction in the building process S.L. (Lieke) Nieman

The Quality Assurance Act for construction in the building process S.L. (Lieke) Nieman

15-08-16 The Quality Assurance Act for construction in the building process S.L. (Lieke) Nieman Construction Management & Engineering July 11, 2016 Table of contents Introduction Problem definition Research question

493 views • 16 slides
The Quality Checker Service Claira Ferreira- NHS Nene and Corby Clinical Commissioning Groups

The Quality Checker Service Claira Ferreira- NHS Nene and Corby Clinical Commissioning Groups

The Quality Checker Service Claira Ferreira- NHS Nene and Corby Clinical Commissioning Groups History behind Quality Checking 2005- Skills for People (Newcastle-Upon-Tyne) 2006- The 11 REACH standards (in supported living) were

372 views • 15 slides
Making Safeguarding Personal 2017/18 Conferences 30 th Jan & 19 March 2018 Making user

Making Safeguarding Personal 2017/18 Conferences 30 th Jan & 19 March 2018 Making user

Making Safeguarding Personal 2017/18 Conferences 30 th Jan & 19 March 2018 Making user involvement effective in Safeguarding Adults Boards Aims of the Conference To identify the key factors that allow users to be involved in Safeguarding

643 views • 49 slides
Some Guidelines on How to Some Guidelines on How to Some Guidelines on How to Deliver a Good

Some Guidelines on How to Some Guidelines on How to Some Guidelines on How to Deliver a Good

Some Guidelines on How to Some Guidelines on How to Some Guidelines on How to Deliver a Good Presentation Deliver a Good Presentation Deliver a Good Presentation Mohammed H. Sqalli Mohammed H. Sqalli Mohammed H. Sqalli King Fahd University

319 views • 16 slides
Spectre/Meltdown at eCG: Rebooting 80k cores Bruno Bompastor Adrian Joian Cloud Reliability Team

Spectre/Meltdown at eCG: Rebooting 80k cores Bruno Bompastor Adrian Joian Cloud Reliability Team

Spectre/Meltdown at eCG: Rebooting 80k cores Bruno Bompastor Adrian Joian Cloud Reliability Team 2018 10 Brands In Multiple Countries NL/DE Datacenters Spectre/Meltdown - Meltdown: melts security boundaries which are normally enforced by

305 views • 26 slides
ID check and proctoring in Zoom at Valborg exams- student question time Lena Peterson &

ID check and proctoring in Zoom at Valborg exams- student question time Lena Peterson &

ID check and proctoring in Zoom at Valborg exams- student question time Lena Peterson & working group LP4 distans 2020-04-29 version 1.1 Working group study period 4 Lena Peterson - management Johanna dman communication

761 views • 13 slides
Compliance Check Process January 2015 AGENDA C OMPLIANCE C HECKS : Introduction

Compliance Check Process January 2015 AGENDA C OMPLIANCE C HECKS : Introduction

Compliance Check Process January 2015 AGENDA C OMPLIANCE C HECKS : Introduction Framework Benefits and Conclusion Tools 2 COMPLIANCE CHECKS - INTRODUCTION W HAT ARE COMPLIANCE CHECKS ? Process to obtain and evaluate

340 views • 8 slides
SpaceWire Remote Terminal Controller Torbjrn Hult 20 July 2005 This document or software is

SpaceWire Remote Terminal Controller Torbjrn Hult 20 July 2005 This document or software is

SpaceWire Remote Terminal Controller Torbjrn Hult 20 July 2005 This document or software is confidential to Saab Ericsson Space and may not: a) be used for any purpose other than for which it was supplied; 1 b) be copied or reproduced in

437 views • 12 slides

More recommend