be230cLecture9

52 rapid analysis of single cell calcium transients

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Unformatted text preview: nces in Vitro to Improve Risk Stratification Using iPS Cells to Fulfill the Promise of Personalized Medicine Patient Perform High Throughput Studies among patient populations with different mutations in different genes to Evaluate for Toxicity, Develop New Therapeutics Etc. 52 Rapid Analysis of Single Cell Calcium Transients Using Kinetic Imaging Cytometer (KIC) Courtesy of Mark Mercola, Vela Laboratories 53 • iPS offers great promise for study of human cardiovascular disesease in vitro • Challenges include • Efficient generation of cells • Studying diseases which are cell autonomous with clear in vitro phenotypes • Neil Chi • Cynthia Perry • Dekker Deacon • Johnathan Grinstein Thank You • Zheng Feng Zhou • Quiming Gong, Matthew Stump • Lei Yang • Ihor Lemischka • Christoph Schaniel • Rocky Kass • Cecile Terronoire, George Wang, Michelle Park Stacey Clegg Aaron Kaplan Jon Lu • • Eric Sobie • Frank Fabris NIH K23, CIRM Gordon Keller, Steve Kattman Dahlia Ruby Can We Make Pacemaker Cells? 40 mV Patch Clamp Analysis 100 ms 500 ms 100 ms 40 mV 40 mV 500 ms 100 ms 100 ms HNK/MinK Expressed at Day 4 of Cardiomyocyte Differentiation Control HNK Expression Non Cardiac 0.82 0.41 5.52 1.05 98.4 0.41 92.6 0.79 2.81 0.7 4.11 1.02 3.32 89.5 3.55 90.9 4.74 0.6 0.5 mINK 0.4 0.3 0.2 0.1 0 Day 1 Day 4 Day 7 Day 9 Day 12 ! Cardiac Day 4 Cardiac Day 6 HNK Cx45 mINK 0.00110 0.020 0.015 0.010 ! 2.81 4.11 0.00095 0.00080 0.005 0.00065 0 0.00050 Hnk+ Hnk- Hnk+ HCN4 Hnk- Cx40 0.0002000 0.00700 0.0001588 0.00525 0.0001175 0.00350 0.0000763 0.00175 0.0000350 0 MinK 89.5 3.55 HNK Hnk+ Hnk- Hnk+ Hnk- HNK Positive Cells Contain Pacemaker Like Cells HNK Positive HNK Negative Notch Signaling Regulates Commitment to Conduction System Development? 4 6C 4 Notch Induced mES Cardiomyocytes 6 C Can We Treat Disease with Stem Cells Does Cell Type Matter? SCID BEIGE mice with induced myocardial infarctions Hemangioblast One Week Post Infarct Cardiac Progenitor MRI EP Study Pathological analysis MRI EP Study Pathological analysis •Mouse intubated •Heart Exposed •LAD visualized, one 8-0 suture placed around it •5 minutes after infarction 10 microliters of cells/ medium directly injected into infarcted area •Total Time 30 min 9.4 Tesla MRI for Evaluation of Murine Cardiac Function ES-Cardiac Cells (notch +) improve Function more than Hemangioblasts (notch -) Notch + Cells became Cardiomyocytes in Vivo Adler et al. Faseb. Nov 2009 Hemangioblasts Form Vascular Cells in Culture Adler et al. Faseb. Nov 2009 Transplanted Hemangioblasts Survive as much as Cardioangioblasts...
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This document was uploaded on 04/09/2014.

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