- Email: [email protected]
Differentiation potential of cardiac stem and progenitor cells in a model of heterotopic heart transplantation
S100
ABSTRACTS / Journal of Molecular and Cellular Cardiology 42 (2007) S88–S101
Sca-1 knockouts mice. In parallel, cardiac non-myocyte cells, previously shown to contain CPCs, were isolated from Sca-1deficient mice and induced to differentiate into cardiomyocytes. The Sca-1-deficient non-myocyte cell population showed a reduced capacity to produce differentiated cardiomyocytes in vitro. All together, these results support a role for Sca-1 and of Sca-1+ cells in maintaining cardiac integrity possibly via the regulation of CPC number and cardiomyocyte differentiation. Keywords: Stem cells; Cardiac remodeling doi:10.1016/j.yjmcc.2007.03.218
Skeletal myoblasts and bone marrow cells enhance contractility of failing cardiomyocytes in co-culture Joon Lee, Urszula Siedlecka, Mark Stagg, Gopal Soppa, Magdi Yacoub, Cesare Terracciano. Harefield Heart Science Centre, NHLI, UK Cell transplantation has been shown to enhance ventricular function in heart failure, but the mechanisms responsible remain unknown. We hypothesized that bone marrow mononuclear cells and skeletal myoblasts cells can influence cardiomyocytes by paracrine mechanisms. Myocardial infarction was induced in adult female Sprague–Dawley rats. Left ventricular ejection fraction after 3 weeks was 35.5 ± 2.0%. Isolated failing left ventricular myocytes were cultured (density 5.2 cells/mm2) on their own (Control) or with either bone marrow mononuclear cells (BM, density 52 cells/mm2) or skeletal myoblasts (SK, density 5.2 cells/mm2). BM and SK were separated from the failing cardiomyocytes by a Transwell™ porous membrane. After 48 h of co-culture with SK or BM cardiomyocyte relaxation speed was increased. The decay time course of the indo-1 transient was also hastened by SK. SK, but not BM, also increased the cardiomyocyte contraction amplitude compared to Control.
Sarcomere shortening (nm) Sarcomere relaxation T50 (ms) Indo-1 decay T50 (ms)
Control
BM
SK
46 ± 6 34 ± 6 22 ± 2
55 ± 1 18 ± 3* 18 ± 2
91±14* 13 ± 2* 13 ± 1*
Data represented as mean ± SEM, *p < 0.05 vs. Control.
We conclude that bone marrow mononuclear cells and skeletal myoblasts enhance cardiomyocyte function in coculture by paracrine mechanisms, which may help explain the functional improvement seen after cell transplantation. Keywords: Heart failure; Cardiomyocytes; Calcium handling doi:10.1016/j.yjmcc.2007.03.219
Cardiac stem cells can be generated in damaged heart from bone marrow-derived cells Lucio Barile, Francesco Cerisoli, Roberto Gaetani, Elvira Forte, Sergio Ottolenghi, Maria C. Magli, E. Messina. Univ. “La Sapienza” Rome, Italy There is now strong evidence that the heart harbours cells with high cardiac muscle regenerative potential. These cells are considered to be cardiac stem cells (CSCs). The origin of adult CSC is completely unknown. Although CSCs express Kit, a membrane receptor that marks several stem cell types, including hematopoietic cells, there is no direct evidence that BM cells can generate functionally pool of CSCs. To investigate whether BM cells can contribute to repopulate the cardiac Kit+ CSCs pool, we transplanted BM cells from a mouse line expressing transgenic green fluorescent protein (GFP) under the control of Kit regulatory elements, into wild type irradiated recipients. After hematological reconstitution (4–5 months) and following a myocardial infarction (MI), cardiac cells were grown in vitro as typical “cardiospheres” (CSs). We next asked whether Kit/ GFP+ CSs were functionally capable of regeneration in vivo. We injected 1 × 105 β-gal labeled CSs-derived cells (CDCs) in the border zone of three wild type mice, immediately after MI. Fluorescent CSs were obtained from the hearts of transplanted and infarcted mice but not from ones that had not undergone MI. Many blue stained CDCs could be found engrafted throughout the infarct region of the mouse heart at 20 days, and a high proportion of them could be identified as differentiated cardiomyocytes and vascular cells within viable myocardium and in scattered islands in the infarct area (but not in the control treated animals). These findings indicate that, at least in conditions of local CSCs depletion bone marrow cells can give rise, after homing into the heart, to cells with properties of resident Kit+ CSCs. Keywords: Stem cells; Heart failure; Bone marrow stromal cells doi:10.1016/j.yjmcc.2007.03.220
Differentiation potential of cardiac stem and progenitor cells in a model of heterotopic heart transplantation T. Zaglia1, S. Bracco1, A. Dedja2, E. Cozzi2, S. Schiaffino1, S. Ausoni1. 1Dept. of Biomedical Sciences, University of Padua, Italy. 2Dept. Med. and Surg. Sciences, University of Padua, Italy Cardiac stem cells were identified in mammalian hearts and they were found to regenerate cardiomyocytes and vessels when injected into an infarcted heart. We used the model of heterotopic heart transplantation to investigate spontaneous potential of cardiac stem and progenitor cells to differentiate properly after tissue damage and remodeling. In a previous study, in which hearts from normal rat donors were heterotopically transplanted into GFP+ transgenic rat hosts, we found abundant extracardiac GFP+ cells that did not contribute to de
ABSTRACTS / Journal of Molecular and Cellular Cardiology 42 (2007) S88–S101
novo cardiogenesis, but generated rare hybrid cardiomyocytes by cell fusion with resident cardiac cells. In the same transplants we recently identified a robust population of GFP− cells expressing markers of stem and mesenchymal cells. Markers of cardiac commitment and neural stem cells identified different populations of cell progenitors. The overall number of these cells increased 5-fold in heart transplants as compared to normal hearts. The majority of these cells did not progress towards maturation, however cardiomyocytes expressing phosphohistone-H3 were occasionally found. A major issue is whether cardiac stem cells can detach from a damaged heart, circulate and from the bloodstream home again into the heart. To address this question the heart of GFP+ transgenic rats was transplanted into a normal host, which had been previously treated with isoproterenol, to induce damage in the native heart. Native hearts, retrieved 15 days after surgery, did not show any significant engraftment by cardiac stem cells mobilized from the heart transplant. Keywords: Stem cells; Differentiation; Cardiac remodeling doi:10.1016/j.yjmcc.2007.03.221
Stem cells recruitment in human right ventricular remodeling A. Angelini, C. Castellani, M. Della Barbera, S. Toffoli, M.A. Padalino, O. Milanesi, G. Stellin, G. Thiene. Cardiovascular Pathology, Pediatric Cardiology and Cardiac Surgery, University of Padua, Padua, Italy Introduction: Aim of the study was to identify the role of cardiac stem cells in right ventricular remodeling of patients affected by Hypoplastic Left Heart Syndrome after Norwood surgical procedures. Methods: Eleven hearts of the affected patients (ages 9 to 365 days) were compared with age-matched controls. Samples taken from the inflow and outflow tracts of the right ventricle were evaluated at histology and immunohistochemistry for stem cells markers. Results: We observed a recruitment of cardiac precursor stem cells (CPSCs) CD117 and CD105 positive, both in the inflow and the outflow tract in operated hearts compared to the controls (CD117 inflow: controls 7.8 ± 3.3, operated hearts 18.09 ± 15.85 cells/field, with p = 0.06; outflow: controls 9 ± 4.73, operated patients 17.81 ± 13.61 cells/field with p = 0.01; CD105 inflow: controls 1.3 ± 0.9, operated patients 9.18 ± 3.8 cells/field, p = 0.07; outflow: controls 1.8 ± 1.6, operated patients 6.2 ± 3.1 cells/field, p=0.1). CPSCs seem to be mainly mobilized by acute
S101
injury (early death patients) rather than by chronic processes (late death patients) since a higher presence of putative cardiac stem cells (CD105) was detected in operated patients. The major fraction of CPSCs were found to be localized near collagen III. Rare cycling cardiac precursor cells, positive both for myosin and Ki67 markers were found. Conclusions: During right ventricle hypertrophy we observed a recruitment of CPSCs, suggesting an active role of endothelial precursor progenitor cells in cardiac remodeling mainly involving the interstitial and endothelial tissues. Keywords: Stem cells; Right ventricular remodeling doi:10.1016/j.yjmcc.2007.03.222
A prognostic indicator for limb amputation after bone marrow mononuclear cells implantation Shuhei Tara, Gen Takagi, Koji Kato, Hitoshi Takano, Ikuyo Takagi, Masahiro Yasutake, Masaaki Miyamoto, Teruo Takano. The First Department of Internal Medicine Nippon Medical School, Japan Objectives: Bone marrow mononuclear cells implantation (BMI) is a promising therapy for severe peripheral artery disease (PAD). However, limb amputation becomes inevitable in some patients, which is hard to predict. The study aimed at finding out a prognostic indicator. Method: In 9 PAD patients with Fontaine class 3–4, ankle– brachial index (ABI), transcutaneous oxygen tension (TcPO2) and tissue blood flow examination using Technecium tetrofosmin scintigraphy (Tc-TF) were performed before and 4 weeks after BMI. We compared changes in those parameters between patients with eventual amputation (group A, n = 3) and those without (group N, n = 6). Results: Prior to BMI, all parameters were comparable between the 2 groups. ABI and Tc-TF did not change significantly in both groups: ABI 0.3 ± 0.4 to 0.3 ± 0.3 in group A, 0.4 ± 0.2 to 0.4 ± 0.09 in group N, NS; Tc-TF 0.4 ± 0.1 to 0.6 ± 0.2 in group A, 0.7 ± 0.1 to 0.9 ± 0.3 in group N, NS, respectively. However, TcPO2 significantly increased from 23.1 ± 7.8 to 40.3 ± 10.8 Torr in group N, whereas it decreased in group A from 12.6 ± 15.3 to 7.6 ± 9.8 Torr, p < 0.05. Conclusion: TcPO2 can be a useful prognostic indicator in severe PAD patients after BMI. Keywords: Angiogenesis; Stem cell doi:10.1016/j.yjmcc.2007.03.223
ABSTRACTS / Journal of Molecular and Cellular Cardiology 42 (2007) S88–S101
Sca-1 knockouts mice. In parallel, cardiac non-myocyte cells, previously shown to contain CPCs, were isolated from Sca-1deficient mice and induced to differentiate into cardiomyocytes. The Sca-1-deficient non-myocyte cell population showed a reduced capacity to produce differentiated cardiomyocytes in vitro. All together, these results support a role for Sca-1 and of Sca-1+ cells in maintaining cardiac integrity possibly via the regulation of CPC number and cardiomyocyte differentiation. Keywords: Stem cells; Cardiac remodeling doi:10.1016/j.yjmcc.2007.03.218
Skeletal myoblasts and bone marrow cells enhance contractility of failing cardiomyocytes in co-culture Joon Lee, Urszula Siedlecka, Mark Stagg, Gopal Soppa, Magdi Yacoub, Cesare Terracciano. Harefield Heart Science Centre, NHLI, UK Cell transplantation has been shown to enhance ventricular function in heart failure, but the mechanisms responsible remain unknown. We hypothesized that bone marrow mononuclear cells and skeletal myoblasts cells can influence cardiomyocytes by paracrine mechanisms. Myocardial infarction was induced in adult female Sprague–Dawley rats. Left ventricular ejection fraction after 3 weeks was 35.5 ± 2.0%. Isolated failing left ventricular myocytes were cultured (density 5.2 cells/mm2) on their own (Control) or with either bone marrow mononuclear cells (BM, density 52 cells/mm2) or skeletal myoblasts (SK, density 5.2 cells/mm2). BM and SK were separated from the failing cardiomyocytes by a Transwell™ porous membrane. After 48 h of co-culture with SK or BM cardiomyocyte relaxation speed was increased. The decay time course of the indo-1 transient was also hastened by SK. SK, but not BM, also increased the cardiomyocyte contraction amplitude compared to Control.
Sarcomere shortening (nm) Sarcomere relaxation T50 (ms) Indo-1 decay T50 (ms)
Control
BM
SK
46 ± 6 34 ± 6 22 ± 2
55 ± 1 18 ± 3* 18 ± 2
91±14* 13 ± 2* 13 ± 1*
Data represented as mean ± SEM, *p < 0.05 vs. Control.
We conclude that bone marrow mononuclear cells and skeletal myoblasts enhance cardiomyocyte function in coculture by paracrine mechanisms, which may help explain the functional improvement seen after cell transplantation. Keywords: Heart failure; Cardiomyocytes; Calcium handling doi:10.1016/j.yjmcc.2007.03.219
Cardiac stem cells can be generated in damaged heart from bone marrow-derived cells Lucio Barile, Francesco Cerisoli, Roberto Gaetani, Elvira Forte, Sergio Ottolenghi, Maria C. Magli, E. Messina. Univ. “La Sapienza” Rome, Italy There is now strong evidence that the heart harbours cells with high cardiac muscle regenerative potential. These cells are considered to be cardiac stem cells (CSCs). The origin of adult CSC is completely unknown. Although CSCs express Kit, a membrane receptor that marks several stem cell types, including hematopoietic cells, there is no direct evidence that BM cells can generate functionally pool of CSCs. To investigate whether BM cells can contribute to repopulate the cardiac Kit+ CSCs pool, we transplanted BM cells from a mouse line expressing transgenic green fluorescent protein (GFP) under the control of Kit regulatory elements, into wild type irradiated recipients. After hematological reconstitution (4–5 months) and following a myocardial infarction (MI), cardiac cells were grown in vitro as typical “cardiospheres” (CSs). We next asked whether Kit/ GFP+ CSs were functionally capable of regeneration in vivo. We injected 1 × 105 β-gal labeled CSs-derived cells (CDCs) in the border zone of three wild type mice, immediately after MI. Fluorescent CSs were obtained from the hearts of transplanted and infarcted mice but not from ones that had not undergone MI. Many blue stained CDCs could be found engrafted throughout the infarct region of the mouse heart at 20 days, and a high proportion of them could be identified as differentiated cardiomyocytes and vascular cells within viable myocardium and in scattered islands in the infarct area (but not in the control treated animals). These findings indicate that, at least in conditions of local CSCs depletion bone marrow cells can give rise, after homing into the heart, to cells with properties of resident Kit+ CSCs. Keywords: Stem cells; Heart failure; Bone marrow stromal cells doi:10.1016/j.yjmcc.2007.03.220
Differentiation potential of cardiac stem and progenitor cells in a model of heterotopic heart transplantation T. Zaglia1, S. Bracco1, A. Dedja2, E. Cozzi2, S. Schiaffino1, S. Ausoni1. 1Dept. of Biomedical Sciences, University of Padua, Italy. 2Dept. Med. and Surg. Sciences, University of Padua, Italy Cardiac stem cells were identified in mammalian hearts and they were found to regenerate cardiomyocytes and vessels when injected into an infarcted heart. We used the model of heterotopic heart transplantation to investigate spontaneous potential of cardiac stem and progenitor cells to differentiate properly after tissue damage and remodeling. In a previous study, in which hearts from normal rat donors were heterotopically transplanted into GFP+ transgenic rat hosts, we found abundant extracardiac GFP+ cells that did not contribute to de
ABSTRACTS / Journal of Molecular and Cellular Cardiology 42 (2007) S88–S101
novo cardiogenesis, but generated rare hybrid cardiomyocytes by cell fusion with resident cardiac cells. In the same transplants we recently identified a robust population of GFP− cells expressing markers of stem and mesenchymal cells. Markers of cardiac commitment and neural stem cells identified different populations of cell progenitors. The overall number of these cells increased 5-fold in heart transplants as compared to normal hearts. The majority of these cells did not progress towards maturation, however cardiomyocytes expressing phosphohistone-H3 were occasionally found. A major issue is whether cardiac stem cells can detach from a damaged heart, circulate and from the bloodstream home again into the heart. To address this question the heart of GFP+ transgenic rats was transplanted into a normal host, which had been previously treated with isoproterenol, to induce damage in the native heart. Native hearts, retrieved 15 days after surgery, did not show any significant engraftment by cardiac stem cells mobilized from the heart transplant. Keywords: Stem cells; Differentiation; Cardiac remodeling doi:10.1016/j.yjmcc.2007.03.221
Stem cells recruitment in human right ventricular remodeling A. Angelini, C. Castellani, M. Della Barbera, S. Toffoli, M.A. Padalino, O. Milanesi, G. Stellin, G. Thiene. Cardiovascular Pathology, Pediatric Cardiology and Cardiac Surgery, University of Padua, Padua, Italy Introduction: Aim of the study was to identify the role of cardiac stem cells in right ventricular remodeling of patients affected by Hypoplastic Left Heart Syndrome after Norwood surgical procedures. Methods: Eleven hearts of the affected patients (ages 9 to 365 days) were compared with age-matched controls. Samples taken from the inflow and outflow tracts of the right ventricle were evaluated at histology and immunohistochemistry for stem cells markers. Results: We observed a recruitment of cardiac precursor stem cells (CPSCs) CD117 and CD105 positive, both in the inflow and the outflow tract in operated hearts compared to the controls (CD117 inflow: controls 7.8 ± 3.3, operated hearts 18.09 ± 15.85 cells/field, with p = 0.06; outflow: controls 9 ± 4.73, operated patients 17.81 ± 13.61 cells/field with p = 0.01; CD105 inflow: controls 1.3 ± 0.9, operated patients 9.18 ± 3.8 cells/field, p = 0.07; outflow: controls 1.8 ± 1.6, operated patients 6.2 ± 3.1 cells/field, p=0.1). CPSCs seem to be mainly mobilized by acute
S101
injury (early death patients) rather than by chronic processes (late death patients) since a higher presence of putative cardiac stem cells (CD105) was detected in operated patients. The major fraction of CPSCs were found to be localized near collagen III. Rare cycling cardiac precursor cells, positive both for myosin and Ki67 markers were found. Conclusions: During right ventricle hypertrophy we observed a recruitment of CPSCs, suggesting an active role of endothelial precursor progenitor cells in cardiac remodeling mainly involving the interstitial and endothelial tissues. Keywords: Stem cells; Right ventricular remodeling doi:10.1016/j.yjmcc.2007.03.222
A prognostic indicator for limb amputation after bone marrow mononuclear cells implantation Shuhei Tara, Gen Takagi, Koji Kato, Hitoshi Takano, Ikuyo Takagi, Masahiro Yasutake, Masaaki Miyamoto, Teruo Takano. The First Department of Internal Medicine Nippon Medical School, Japan Objectives: Bone marrow mononuclear cells implantation (BMI) is a promising therapy for severe peripheral artery disease (PAD). However, limb amputation becomes inevitable in some patients, which is hard to predict. The study aimed at finding out a prognostic indicator. Method: In 9 PAD patients with Fontaine class 3–4, ankle– brachial index (ABI), transcutaneous oxygen tension (TcPO2) and tissue blood flow examination using Technecium tetrofosmin scintigraphy (Tc-TF) were performed before and 4 weeks after BMI. We compared changes in those parameters between patients with eventual amputation (group A, n = 3) and those without (group N, n = 6). Results: Prior to BMI, all parameters were comparable between the 2 groups. ABI and Tc-TF did not change significantly in both groups: ABI 0.3 ± 0.4 to 0.3 ± 0.3 in group A, 0.4 ± 0.2 to 0.4 ± 0.09 in group N, NS; Tc-TF 0.4 ± 0.1 to 0.6 ± 0.2 in group A, 0.7 ± 0.1 to 0.9 ± 0.3 in group N, NS, respectively. However, TcPO2 significantly increased from 23.1 ± 7.8 to 40.3 ± 10.8 Torr in group N, whereas it decreased in group A from 12.6 ± 15.3 to 7.6 ± 9.8 Torr, p < 0.05. Conclusion: TcPO2 can be a useful prognostic indicator in severe PAD patients after BMI. Keywords: Angiogenesis; Stem cell doi:10.1016/j.yjmcc.2007.03.223