Il trapianto cellulare come alternativa al trapianto d'organo: studio dei processi di crescita e differenziamento delle cellule staminali orientate verso fenotipi cardiovascolari
Il trapianto cellulare come alternativa al trapianto d'organo: studio dei processi di crescita e differenziamento delle cellule staminali orientate verso fenotipi cardiovascolari
concentration of NO released in the culture medium of control stem cells will first be estimated measuring nitrite/nitrate either by highly sensible DAN methods or by the use oh HPLC. In the case in which these methods will not give the satisfactory results, measurement of nitrite/nitrate by amperometric gas sensor will also be performed;
constitutive NOS activity in control stem cells will be estimated as described in Methods;
effect of NO-donors such as S-nitroslaminopenicilline, sodium nitroprusside and NORs, shock wave and a number of compounds extracted from plants such as polyphenols and terpenes either on the concentration of NO in the culture medium or on the NOS activity in the stem cells will be examined;
effect of combined use of NO-donors, shock wave and natural compounds on the concentration of NO in the culture medium of stem cells or on the NOS activity in the stem cells will also be examined;
differentiation state of treated stem cells will be analysed by cytofluorimetric method and/or immunocytological method;
treated stem cell proliferation will be analysed by measuring the [3H]L-thymidine incorporation;
most promising conditions to achieve more stem cells with distinct differentiation state will be chosen among all tested conditions to be utilised in the phase 2.
PHASE 2
stem cells prepared from bone marrow of the patients will be treated under conditions established in the phase. Cellular proliferation and stabilisation will be analysed as in phase 1;
before transplanting stem cells, cardiac activity of patients will clinically and instrumentally be evaluated;
suspension of enriched stem cells will be inoculated by cardiac catheter to perilesional artery of the patients;
after transplanting stem cells, clinical-and instrumental cardiac activity of patients will carefully be evaluated;
recovery of the cardiac activity of patients will clinically and instrumentally be evaluated.