Closed-chest cell injections into mouse myocardium guided by high-resolution echocardiography.

The mouse is an important model for the development of therapeutic stem cell/bone marrow cell implantation to treat ischemic myocardium. However, its small heart size hampers accurate implantation into the left ventricular (LV) wall. Precise injections have required surgical visualization of the heart, which is subject to complications and is impractical for delayed or repeated injections. Furthermore, the thickness of the myocardium is comparable to the length of a needle bevel, so surgical exposure does not prevent inadvertent injection into the LV cavity. We describe the use of high-resolution echocardiography to guide nonsurgical injections accurately into the mouse myocardial wall. We optimized this system by using a mixture of ultrasound contrast and fluorescent microspheres injected into the myocardium, which enabled us to interpret the ultrasound image of the needle during injection. Quantitative dye injection studies demonstrated that guided closed-chest injections and open-chest injections deliver comparable amounts of injectate to the myocardium. We successfully used this system in a mouse myocardial infarction model to target the injection of labeled cells to a region adjacent to the infarct. Intentional injection of tracer into the LV cavity resulted in a small accumulation in the myocardium, suggesting that non-guided cell injections into mouse hearts may appear to be successful even if the majority of the injectate is lost in the chamber. The use of this system will allow more precise cellular implantation into the mouse myocardium by accurately guiding injections to desired locations, confirming successful implantation of cells, in a clinically relevant time frame.

[1]  D. Torella,et al.  Bone Marrow Cells Differentiate in Cardiac Cell Lineages After Infarction Independently of Cell Fusion , 2004, Circulation research.

[2]  Y. Kan,et al.  Adeno-Associated Viral Vector Delivered Cardiac-Specific and Hypoxia-Inducible VEGF Expression in the Ischemic Mouse Hearts , 2022 .

[3]  C. Nienaber,et al.  CABG and bone marrow stem cell transplantation after myocardial infarction. , 2004, The Thoracic and cardiovascular surgeon.

[4]  M. Gnecchi,et al.  Molecular and cell-based therapies for protection, rescue, and repair of ischemic myocardium: reasons for cautious optimism. , 2004, Circulation.

[5]  I. Weissman,et al.  Haematopoietic stem cells adopt mature haematopoietic fates in ischaemic myocardium , 2004, Nature.

[6]  David A. Williams,et al.  Haematopoietic stem cells do not transdifferentiate into cardiac myocytes in myocardial infarcts , 2004, Nature.

[7]  C. Murry,et al.  Evidence for Fusion Between Cardiac and Skeletal Muscle Cells , 2004, Circulation research.

[8]  J. Leor,et al.  Interventional magnetic resonance imaging for guiding gene and cell transfer in the heart , 2003, Heart.

[9]  Klaus Pfeffer,et al.  Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes , 2003, Nature.

[10]  J. Ingwall,et al.  Mesenchymal stem cells modified with Akt prevent remodeling and restore performance of infarcted hearts , 2003, Nature Medicine.

[11]  E. Foster,et al.  The α1A/C- and α1B-adrenergic receptors are required for physiological cardiac hypertrophy in the double-knockout mouse , 2003 .

[12]  H. Blau,et al.  Localized arteriole formation directly adjacent to the site of VEGF-induced angiogenesis in muscle. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[13]  D. Turnbull,et al.  Onset of Cardiac Function During Early Mouse Embryogenesis Coincides With Entry of Primitive Erythroblasts Into the Embryo Proper , 2003, Circulation research.

[14]  Hung-Fat Tse,et al.  Angiogenesis in ischaemic myocardium by intramyocardial autologous bone marrow mononuclear cell implantation , 2003, The Lancet.

[15]  Bernd Westphal,et al.  Autologous bone-marrow stem-cell transplantation for myocardial regeneration , 2003, The Lancet.

[16]  P. Wernet,et al.  Repair of Infarcted Myocardium by Autologous Intracoronary Mononuclear Bone Marrow Cell Transplantation in Humans , 2002, Circulation.

[17]  R. Klein.  Adeno-associated virus vectorology and gene therapy applications , 2002 .

[18]  Orlando Aristizábal,et al.  Spatial velocity profile in mouse embryonic aorta and Doppler-derived volumetric flow: a preliminary model. , 2002, American journal of physiology. Heart and circulatory physiology.

[19]  E. Chérin,et al.  A new ultrasound instrument for in vivo microimaging of mice. , 2002, Ultrasound in medicine & biology.

[20]  J. Isner.  Myocardial gene therapy , 2002, Nature.

[21]  H. Blau,et al.  Gene Delivery to Muscle , 2001, Current protocols in human genetics.

[22]  H. Blau,et al.  Myoblast-mediated gene transfer for therapeutic angiogenesis. , 2002, Methods in enzymology.

[23]  W Grossman,et al.  LV systolic performance improves with development of hypertrophy after transverse aortic constriction in mice. , 2001, American journal of physiology. Heart and circulatory physiology.

[24]  David M. Bodine,et al.  Bone marrow cells regenerate infarcted myocardium , 2001, Nature.

[25]  H. Blau,et al.  VEGF gene delivery to myocardium: deleterious effects of unregulated expression. , 2000, Circulation.

[26]  H. Blau,et al.  Angiogenesis monitored by perfusion with a space-filling microbead suspension. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[27]  Thomas N. Sato,et al.  Leakage-resistant blood vessels in mice transgenically overexpressing angiopoietin-1. , 1999, Science.

[28]  Tomoko Nakanishi,et al.  ‘Green mice’ as a source of ubiquitous green cells , 1997, FEBS letters.

[29]  N. Dracopoli,et al.  Current protocols in human genetics , 1994 .

[30]  H. Blau,et al.  Primary mouse myoblast purification, characterization, and transplantation for cell-mediated gene therapy , 1994, The Journal of cell biology.