Manual of research techniques in cardiovascular medicine / edited by Hossein, Ardehali, Roberto Bolli, Douglas W. Losordo.

Format:

Book

Contributor:

Ardehali, Hossein.

Bolli, Roberto.

Losordo, Douglas W.

Language:

English

Subjects (All):

Angioplasty.

Coronary heart disease--Treatment.

Coronary heart disease.

Tachycardia--Treatment.

Tachycardia.

Transluminal angioplasty.

Physical Description:

1 online resource (xx, 451 pages) : illustrations

Edition:

1st ed.

Place of Publication:

Chichester, West Sussex ; Hoboken : Wiley, 2014.

Language Note:

English.

System Details:

Mode of access: World Wide Web.

Summary:

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.

Contents:

Cover

Dedication

Title page

Copyright page

Contents

List of Contributors

Preface

About the Companion Website

Part 1: Electrophysiology

1: Measurement of calcium transient ex vivo

Introduction

Preparation and fluorescent indicator loading

Optical setup

Calibration

Analysis and interpretation

Set-by-step procedure

References

2: Confocal imaging of intracellular calcium cycling in isolated cardiac myocytes

Materials and methods

Chamber, electrical stimulation, and superfusate delivery apparatus

Confocal microscope system

Cell loading of fluorescent Ca2+ indicators

Experimental approach

Data analyses

Weaknesses and strengths of the method and alternative approaches

Conclusions

3: Generating a large animal model of persistent atrial fibrillation

Insights from tachypacing-induced AF models

Protocol

Generating a tachypacing-induced AF model

Pacing protocols

Ventricular rate during AF: to ablate or not to ablate

Strengths and weaknesses of the model

Complications and alternative approaches to study AF

Potential complications

Alternative AF models

Conclusion

Acknowledgement

4: Confocal imaging of intracellular calcium cycling in the intact heart

Measurement of intracellular Ca2+ transients

Principles and features of confocal microscopy

Procedure

Materials and equipment

Experimental setup

Limitations, potential pitfalls and alternative approaches

Additional application: use of confocal imaging to determine efficacy of stem cell integration into host myocardium

5: Recording and measurement of action potentials

Theory of AP recording

Recording amplifier.

Choices to make before an experiment

AP recording procedure

Frequent problems

AP analysis

Alternative approaches and extensions

Weaknesses and strengths

6: Patch-clamp recordings from isolated cardiac myocytes

Theory of patch-clamp recording

Cell-attached configuration

Whole-cell recording configuration

Inside-out excised patch configuration

Outside-out excised patch configuration

Instrumentation for patch-clamp recordings

Experimental aspects of patch-clamp recordings

Gigaseal formation

Whole-cell recordings

Whole-cell voltage-clamp protocols, data acquisition, and analysis

Conclusions and future directions

7: Optical mapping of the heart

Protocols

Experimental set-up

Tissue preparations (Figure 7.2)

Staining and image acquisition

Data processing and analysis

Representative results

Alternative approaches

Weaknesses and strengths of method

Part 2: Isolation and Maintenance of Primary Stem Cells

8: Isolation of colony-forming endothelial progenitor cells

Materials

Laboratory reagents

Laboratory equipment

Methods

MNC isolation from fresh PB or UCB

EPC-CFA of bulk cell populations in PB, GmPB, BM, or UCB

Optional-1: EPC-CFA of single CD34+ or CD133+ cells in PB, GmPB, BM, or UCB

Optional-2: HELIC assay of single CD34+ or CD133+ cells as cell fate assay of HSC

Application

Acknowledgements

Competing interests statement

9: Cardiac resident stem cells

Preparation of basic solutions and culture medium

Enzymatic digestion of the mouse heart

Separation of small cells

Sorting c-kit-positive cells using FACS

Weaknesses and strength of the method.

Conclusion

10: Cardiospheres

Preparation for explant creation

Explant creation

Preparation for cardiosphere formation

Cardiosphere formation

Weaknesses and strengths of the method

11: Mesenchymal stem cells

Isolation of human MSCs from BM aspirates

12: Generation and differentiation of human iPS cells

Protocol I: Generation of hiPSC from fibroblasts

Production of the VSV-G Lentivirus (6 days)

Reprogramming of human fibroblasts (∼14 days)

Plasmids and media

Titration of the virus

Protocol II: differentiation hiPSCs to myocytes

Future perspectives

13: Isolation of neonatal and adult rat cardiomyocytes

Isolation of neonatal rat ventricular cardiomyocytes

Isolation of adult rat cardiomyocytes

14: Isolation and culture of vascular smooth muscle cells

Reagents

Equipment

Reagent setup

Preparation and equipment setup

Explant method

Strengths and weaknesses of method

15: Isolation and culture of cardiac endothelial cells

Protocol for isolation of mouse cardiac endothelial cells

Materials and preparations

Procedures

Important notes

A novel protocol of endothelial cell isolation

16: Isolation and culture of cardiac fibroblasts

Cardiac fibroblasts isolation protocol

Materials and preparations.

Procedures for isolation of mouse cardiac fibroblasts

Cell propagation and maintenance

Isolation of cardiac ventricular fibroblasts from rats

17: Murine bone marrow transplantation model

Animal husbandry

18: In vitro differentiation and expansion of vascular endothelial cells derived from mouse embryonic stem cells

Tissue culture

ES-cell culture (see Notes 1 and 2)

Differentiation of ES-cells to Flk-1+ progenitors

Purification of Flk-1+ cells

Expansion/differentiation of Flk-1+ cells

Notes

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

19: Coronary ligation

Acknowledgments

20: Transverse aortic constriction: a model to study heart failure in small animals

Material

Preparation of the operative field

Anesthesia, intubation, and ventilation

Aortic banding (constriction of transverse aorta)

Postoperative care

Limitations

Confirmation of constriction

Expected outcomes

21: Pharmacological models of hypertrophy and failure

Overview

Strengths and weaknesses of the method

22: Hindlimb ischemia

Induction of anesthesia

Hair removal prior to surgery

Surgery preparation

Femoral artery ligation

Laser Doppler

Alternative methods.

Common considerations

23: The Langendorff preparation

Cannulation of the aorta

Perfusion solution constituents, thermoregulation, and mode of delivery

Recording of left ventricular pressure, ECG, and coronary flow

24: Myocarditis and other immunological models of cardiac disease

Induction of EAM

Strength and weaknesses of the method

25: Models of pacing-induced heart failure

Rapid pacing models - protocols

Functional and hemodynamic expectations

Neurohormonal aspects and expectations

Cellular and biochemical considerations to chronic rapid pacing

Matrix remodeling and chronic rapid pacing

Electrophysiology and arrhythmogenesis in chronic rapid pacing

Chronic rapid pacing-induced heart failure: the issue of reversibility

Chronic pacing model for therapeutic research and development: technical considerations and limitations

Summary

26: Porcine myocardial ischemia models

Anesthetized pig model with an angioplasty balloon-induced occlusion

Conscious pig model with an extravascular balloon-induced occlusion

27: Angiogenesis assays

In vivo Matrigel plug assay

Preparation of reagents and animals

Reagents for quantification of angiogenesis

Matrigel mixtures and injections

Matrigel harvest and analysis

Mouse corneal micropocket angiogenesis assay

Preparation of controlled slow-release pellet.

Surgical procedure.

Notes:

Bibliographic Level Mode of Issuance: Monograph.

Includes bibliographical references and index.

Description based on online resource; title from PDF title page (ebrary, viewed January 16, 2014).

ISBN:

9781118495131

1118495136

9781118495148

1118495144

9781118495162

1118495160

OCLC:

868925744