Journal of Cardiovascular Medicine and Cardiology eertechz Citation: Francesconi A, Paparella G, Frossi B, Del Terra E, Ambesi Impiombato FS, et al. (2015) C-Kit Positive Cells from Failing Human Hearts: Role of Culturing Media on Cardiomyogenic Potentials. J Cardiovasc Med Cardiol 2(2): 021-028. 021 Abstract Background: The possibility of culturing heart cells in order to regenerate damaged tissue is a challenging problem. Recent observations have demonstrated the possibility of isolating and expanding resident cardiac stem cells, which could favor regeneration and functional improvement of the myocardial tissue. Aims: To investigate two different culturing media: one promoting c-kit cells’ growth and the other promoting differentiation in cardiac muscle cells. Methods: We obtained primary cultures from left ventricle myocardial tissues of 10 human hearts of patients with end-stage heart failure who received heart transplantation. Cells were irst cultured in a medium containing high serum and low calcium/magnesium (Ca 2+ /Mg 2+ ) to promote cell growth (medium A). Than they were cultured in another medium that contained lower serum concentration and a variety of different factors in order to induce cell differentiation (medium B). The presence of c-kit, speciic for stem cells, α-sarcomeric actin (SA), speciic for skeletal and cardiac muscle cells, and α-smooth muscle actin (SMA), speciic for smooth muscle cells was studied by immune-cytochemical analysis. Results: A high percentage of c-kit+, SMA-, SA- cells was observed in medium A; in medium B with lower serum and higher Ca 2+ /Mg 2+ concentrations cells became c-kit-, AML+, SA+. In medium A, 78% of the cells were positive for c-kit. After culturing the same cell populations in medium B with lower serum and higher Ca 2+ /Mg 2+ concentrations, the percentage of c-kit positive cells decreased to 21% while the cells positive for SMA and for SA increased respectively from 28 to 82% and from 0 to 59%. Conclusions: Our results conirm the presence of a high percentage of c-kit positive cells in failing human myocardium and, for the irst time, suggest a key role of calcium/magnesium concentration in promoting both c-kit cells’ growth and their differentiation in human cardiac muscle. human heart tissue and successfully expanded them in culture as cardiospheres (CS) [14]. hey have also obtained cell diferentiation of the Cardiospheres Derived Cells (CDC) and in-vivo engratment in post-infarcted tissue [15]. hese results seem to give a great opportunity for making autologous transplants in patients with reduced functional myocardial tissue. Although this last possibility seems very promising there are some diiculties in culturing diferentiated cells. In the mentioned experiments, co-cultures with mouse myocytes seemed to give the right commitment to undiferentiated cells to become diferentiated myocytes. he latter were characterized by autonomous electrical capacity and successful engratment both morphological and functional [15]. Aiming to develop diferentiated myocytes we have cultured human myocardial tissue using two media: one to promote cell growth rather than cell diferentiation using high serum concentration and low Ca 2+ /Mg 2+ (medium A) and the other to promote cell diferentiation using a medium with lower serum concentration and high Ca 2+ /Mg 2+ (medium B). Introduction Culturing heart cells is still a challenging problem due to the absence of suitable cell lines enabling easy gene expression manipulation and notion [1]. In the last decade many groups have been culturing human heart cells with the intent of reproducing feasible myocardial tissue. he possibility of culturing stem cells has given a great impulse to this attempt and many advances have been made in two main directions: culturing embryonic [2,3] and adult stem cells. Adult-derived cardiomyocytes have been obtained from extra- cardiac stem cells found in peripheral tissues such as bone marrow [4,5], whole blood [6] and muscle tissue [7]. Progresses have been made also in the sense of in vivo engratment of the obtained diferentiated cells into myocardial tissue [8,9] despite diiculties in making functional engratments. It has been recently demonstrated that cardiac-stem cells could be “circulating” cells similar to those found in extra-cardiac tissue [10], as well as local “residential” cells constitutionally present [11-13]. Messina and coworkers have identiied multipotent cells in adult Research Article C-Kit Positive Cells from Failing Human Hearts: Role of Culturing Media on Cardiomyogenic Potentials Anna Francesconi 1 , Gaetano Paparella 2 *, Barbara Frossi 3 , Elisa Del Terra 4 , Francesco Saverio Ambesi Impiombato 4 and Francesco Curcio 4 1 Heart Rhythm Management Center, Vrije Universiteit van Brussel, Laarbeeklaan 101, 1090, Brussel, Belgium 2 Heart Rhythm Division, Brussel Heart Center, Clinique S. Jean, Boulevard du Jardin Botanique, 32 - 1000, Brussel, Belgium 3 Department of Biomedical Sciences and Technologies, Immunology Section, University of Udine, P. le S. Maria della Misericordia, 1 – 33100, Udine, Italy 4 Department of Pathology and Experimental Medicine, University of Udine, P.le S. Maria della Misericordia, 1 – 33100, Udine, Italy Dates: Received: October 16, 2014; Accepted: March 04, 2015; Published: March 07, 2015 *Corresponding author: Gaetano Paparella, MD, Department of Cardiology - Brussel Heart Center - Clinique S. Jean - Boulevard du Jardin Botanique, 32 - 1000 - Bruxelles, Brussel - Belgium, Tel : 0032 02 2219111; E-mail: www.peertechz.com Keywords: Heart failure; Cell culture; C-kit; Differentiation