Manual of Research Techniques in CardiovascularMedicine
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More About This Title Manual of Research Techniques in CardiovascularMedicine

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While some research methods or techniques are applicable in several areas of medicine, research in cardiovascular diseases requires knowledge of an increasing array of procedures, techniques and measurements that are highly specialized and unique to this area of investigation. Edited by senior clinical investigators who are recognized leaders in cardiovascular medicine worldwide, this book provides readers with a comprehensive, practical “how-to-do-it” review of best-practice techniques for cardiovascular research.

English

Edited by

Hossein Ardehali MD, PhD, FAHA, Associate Professor of Medicine, Molecular Pharmacology and Biological
Chemistry
Feinberg Cardiovascular Research Institute
Northwestern University
Feinberg School of
Medicine, Chicago, IL USA

Roberto Bolli MD
Professor of Medicine Chief, Division of Cardiovascular Medicine
Director, Institute of Molecular Cardiology
University of Louisville
Louisville, KY USA

Douglas W. Losordo MD
Vice President New Therapeutic Development
Baxter Healthcare Corporation
Deerfield, IL USA

 

English

List of Contributors ix

Preface xix

About the Companion Website xx

Part 1 Electrophysiology

1 Measurement of calcium transient ex vivo 3
Kenneth R. Laurita and Bradley N. Plummer

2 Confocal imaging of intracellular calcium cycling in isolated cardiac myocytes 12
Søren Grubb, J. Andrew Wasserstrom, and Gary L. Aistrup

3 Generating a large animal model of persistent atrial fi brillation 20
Raphaël P. Martins and José Jalife

4 Confocal imaging of intracellular calcium cycling in the intact heart 32
Neha Singh, Manvinder Kumar, James E. Kelly, Gary L. Aistrup, and J. Andrew Wasserstrom

5 Recording and measurement of action potentials 41
Kenneth S. Ginsburg and Donald M. Bers

6 Patch-clamp recordings from isolated cardiac myocytes 50
Kai-Chien Yang, Wei Wang, and Jeanne M. Nerbonne

7 Optical mapping of the heart 60
Di Lang, Sarah Gutbrod, Jacob Laughner, and Igor R. Efi mov

Part 2 Isolation and Maintenance of Primary Stem Cells

8 Isolation of colony-forming endothelial progenitor cells 71
Haruchika Masuda and Takayuki Asahara

9 Cardiac resident stem cells 87
João Ferreira-Martins, Fumihiro Sanada, and Marcello Rota

10 Cardiospheres 95
Rachel Ruckdeschel Smith

11 Mesenchymal stem cells 104
Jose S. Da Silva and Joshua M. Hare

12 Generation and differentiation of human iPS cells 110
Sebastian Diecke, Lei Ye, Sophia Zhang, and Jianyi Zhang

13 Isolation of neonatal and adult rat cardiomyocytes 117
Md. Abdur Razzaque and Jeffrey Robbins

14 Isolation and culture of vascular smooth muscle cells 125
Milton Hamblin, Lin Chang, and Y. Eugene Chen

15 Isolation and culture of cardiac endothelial cells 131
Asish K. Ghosh, Joseph W. Covington, and Douglas E. Vaughan

16 Isolation and culture of cardiac fi broblasts 140
Asish K. Ghosh, Joseph W. Covington and Douglas E. Vaughan

17 Murine bone marrow transplantation model 146
Prasanna Krishnamurthy, Suresh Kumar Verma, and Raj Kishore

18 In vitro differentiation and expansion of vascular endothelial cells derived from mouse embryonic stem cells 149
Anees Fatima, Carey Nassano-Miller, and Tsutomu Kume

Part 3 Manipulation of the Heart and Vessels in Vivo and ex Vivo

19 Coronary ligation 157
Alexander R. Mackie and Hossein Ardehali

20 Transverse aortic constriction: a model to study heart failure in small animals 164
Suresh Kumar Verma, Prasanna Krishnamurthy, and Raj Kishore

21 Pharmacological models of hypertrophy and failure 170
Angela C. deAlmeida, Tariq Hamid, and Sumanth D. Prabhu

22 Hindlimb ischemia 177
Jerry C. Lee, Ngan F. Huang, and John P. Cooke

23 The Langendorff preparation 187
Hugh Clements-Jewery and Michael J. Curtis

24 Myocarditis and other immunological models of cardiac disease 197
Daniela C¡ iháková and Noel R. Rose

25 Models of pacing-induced heart failure 203
James A. Shuman, Rupak Mukherjee, and Francis G. Spinale

26 Porcine myocardial ischemia models 215
Xian-Liang Tang and Roberto Bolli

27 Angiogenesis assays 225
Susmita Sahoo and Douglas W. Losordo

28 Immunohistochemical analysis of cardiac tissue 232
Barbara Ogórek, Donato Cappetta, and Jan Kajstura

29 A murine model of cardiac arrest by exsanguination 237
Guangming Cheng, Yiru Guo, Harold K. Elias, Carrie M. Quinn, Arash Davani, Yanjuan Yang, Magdy Girgis, Roberto Bolli, and Buddhadeb Dawn

Part 4 Small Animal Imaging

30 Blood pressure, telemetry, and vascular measurements in the rodent model 245
Robert S. Danziger

31 The setting: imaging conscious, sedated, or anesthetized rodents 250
Gene H. Kim and Roberto M. Lang

32 Echocardiography: standard techniques (M-mode, two-dimensional imaging, and Doppler) 254
Gene H. Kim, Lauren Beussink-Nelson, Sanjiv J. Shah, and Roberto M. Lang

33 Echocardiography: advanced techniques (tissue Doppler, speckle tracking, and three-dimensional imaging) 275
Lauren Beussink-Nelson, Gene H. Kim, Roberto M. Lang, and Sanjiv J. Shah

34 In vivo tomographic cardiac imaging: positron emission tomography and magnetic resonance imaging 287
Bruno C. Huber, Patricia K. Nguyen, and Joseph C. Wu

35 In vivo hemodynamics 295
Alexander R. Mackie, Kyle K. Henderson, Sol Misener, and Hossein Ardehali

Part 5 Metabolism, Mitochondria, and Cell Death

36 Fractionation of cardiomyocytes and isolation of mitochondria 307
Christopher P. Baines

37 Assessment of glucose and fatty acid metabolism ex vivo 313
Darrell D. Belke and E. Dale Abel

38 Quantification and characterization of atherosclerotic lesions in mice 320
Abhinav Agarwal, Millicent G. Winner, Srinivas D. Sithu, and Sanjay Srivastava

39 Assessment of cell death in the heart 332
Russell S. Whelan, Klitos Konstantinidis, and Richard N. Kitsis

40 Assessment of mitochondrial function in isolated cells 343
Amy K. Rines and Hossein Ardehali

41 Multinuclear NMR spectroscopy of myocardial energetics and substrate utilization in isolated perfused mouse hearts 351
Stephen C. Kolwicz, Jr. and Rong Tian

42 Measurement of reactive oxygen species in cardiovascular disease 359
Mahmood Khan, Fatemat Hassan, Sashwati Roy, and Chandan K. Sen

43 Assessing autophagy 371
Roberta A. Gottlieb

44 Assessment of cardiomyocyte size 378
A. Martin Gerdes and Alessandro Pingitore

Part 6 Manipulation of Gene Expression in Vitro and in Vivo

45 Generation of Cre-loxP mouse models for conditional knockout and overexpression of genes in various heart cells 389
Marisa Z. Jackson and Warren G. Tourtellotte

46 Modulation of myocardial genes via use of adenoviral vectors and RNA interference approaches 397
Qianhong Li

47 Overexpression and downregulation of proteins in vitro 407
Marina Bayeva and Hossein Ardehali

48 In vivo microRNA studies 416
Eva van Rooij

Part 7 Model Systems

49 Vascular and cardiac studies in zebrafi sh 425
Hans-Georg Simon, Molly Ahrens, and Brandon Holtrup

50 Vascular and cardiac studies in Drosophila 432
Lin Yu, Joseph P. Daniels, and Matthew J. Wolf

Index 441

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