Introduction Skeletal Myoblasts cells diferentiation

1. Simultaneous transplantation of co-cultured mesenchymal stem cells and skeletal myoblasts improves ventricular function in Chagas disease • Luiz César Guarita-Souza MD – PhD • PUCPR - Experimental Laboratory of Cell Culture Institute of • Biological and Health SciencesCuritiba-Pr-Brazil

2. Introduction Skeletal Myoblasts cells diferentiation Skeletal Fibers Myocardial Scar Guarita-Souza LC, Int J Cardiol. 2006 Aug 28;111(3):423-9

3. Introduction Mesenchymal Stem cells diferentiation Neoangiogenesis Guarita-Souza LC, Transplant Proc. 2006Jul-Aug;38(6):1953-4

4. Introduction Myocardial Cell Therapy Mesenchymal Stem Cells Skeletal Myoblast + Angio-muscular Regeneration

5. Introduction Myocardial Cell Therapy • The Association of Myoblast and Mesenchymal Stem Cells Transplantation, on Post Infarct Ventricular Dysfunction • Guarita-Souza LC; Carvalho KAT, Rebelatto C; Senegaglia A, Myiague N; Furuta M; Francisco J, Olandowski M, Hansen P. Arq. Bras. Cardiol.2004, OctOct;83(4):294-9

6. Introduction • Cell Therapy • Cell Therapy in Myocardial Infarction • Segmentar • Myocardial Scar and Ischemic • BothCell Therapy in a Cardiomyopathy non-ischemic ?

7. Objective Evaluated the histological and functional effects of simultaneous autologous transplantation of co-cultured mesenchymal and skeletal myoblasts cells in an experimental model of dilated cardiomyopathy due to Chagas disease.

8. Introduction Chagas Disease Fisiopathology • Transmission • Triatoma Infestans: • 5 to 14 days:sains and symptoms • Transfusional: • 30 to 40 days symptoms • Chronic form: after 10 years Immunologic Activity: • Humoral and Cellular Immunity Triatoma Infestans “Barbeiro”

9. Introduction Chagas Disease Fisiopathology • Forms of Disease • Acute • Indeterminated 60% • Cardiac 30% after 10-20y • Digestive • Both • Congenital Trypanossoma Cruzi Myocardial Infection

10. Introduction Chagas Disease Pathology • Chronic myocardite • Chronic inflamatory process • diffuse fibrosis • Impairment of microcirculation • Ischemic myocardium Chronic inflamatory process Myocardial scar HE, 200x

11. Methods Protocol of Research Muscular Biopsy Histology T Cruzi 150.000 forms Animals inoculation 14 of days cultive 8 M 7 days TX 1 Month Bone Marrow Aspiartion Eco 1M Eco Baseline

12. Cell Therapy in MyocardiumProtocol of Cell Culture

13. Material and Method • 80 animals were inoculated - 200 gramas • 21.25%- 17 animals included - FE < 37% • Co-culture group: 9 animals mesenchymal stem cells skeletal myoblast cells 0.30 ml - 5,4x106 8,0x106cells • Control group : 8 animals saline - 0.30 ml

14. Material and MethodEchocardiographic study • Bi-Dimensional Echocardiographic study HP/SONOS 5500- Linear Transduction of 15MHz • After eigth months of inoculation • One month after transplantation • Parameters Analysis • LVEDV (ml) • LVESV (ml) • LVEF (%)

15. ResultsEchocardiographic study

16. Results LVEF(%) (*) Paired Student’s test p<0,05

17. Results LVESV (ml) (*) Paired Student’s test p<0,05

18. Results LVEDV (ml) (*) Paired Student’s test p<0,05

19. ResultsPathology

20. Results Phatology Treat group Control group HE, 200X

21. Results Phatology 200X 400X 400X Treat group Gomery Trichrome

22. Results Phatology / Immunostained Control Treat group Fast Miosin, 40x

23. Conclusion • The co-transplant of mesenchymal stem cell and skeletal myoblasts cell is functionally effective in the Chagas disease ventricular dysfunction.