C-Kit Positive Cells from Failing Human Hearts: Role of Culturing Media on Cardiomyogenic Potentials
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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 first cultured in a medium containing high serum and low calcium/magnesium (Ca2+ /Mg2+) 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, specific for stem cells, α-sarcomeric actin (SA), specific for skeletal and cardiac muscle cells, and α-smooth muscle actin (SMA), specific 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 Ca2+/Mg2+ 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 Ca2+ /Mg2+ 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 confirm the presence of a high percentage of c-kit positive cells in failing human myocardium and, for the first time, suggest a key role of calcium/magnesium concentration in promoting both c-kit cells' growth and their differentiation in human cardiac muscle.
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Copyright (c) 2015 Francesconi A, et al.
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Germani A, Di RG, Limana F, Martelli F (2007) Capogrossi MC. Molecular mechanisms of cardiomyocyte regeneration and therapeutic outlook. Trends Mol Med 13: 125-133. DOI: https://doi.org/10.1016/j.molmed.2007.01.002
Tomita Y, Matsumura K, Wakamatsu Y (2005) Cardiac neural crest cells contribute to the dormant multipotent stem cell in the mammalian heart. J Cell Biol 170: 1135-1146. DOI: https://doi.org/10.1083/jcb.200504061
Fukuda K (2005) Progress in myocardial regeneration and cell transplantation. Circ J 69: 1431-1446. DOI: https://doi.org/10.1253/circj.69.1431
Ferrari G, Cusella-De AG, Coletta M (1998)Muscle regeneration by bone marrow-derived myogenic progenitors. Science 279: 1528-1530. DOI: https://doi.org/10.1126/science.279.5356.1528
Orlic D, Kajstura J, Chimenti S (2001)Bone marrow cells regenerate infarcted myocardium. Nature 410: 701-705. DOI: https://doi.org/10.1038/35070587
Asahara T, Murohara T, Sullivan A (1997)Isolation of putative progenitor endothelial cells for angiogenesis. Science 275: 964-967. DOI: https://doi.org/10.1126/science.275.5302.964
Menasche P (2008)Skeletal myoblasts and cardiac repair. J Mol Cell Cardiol45:545-553. DOI: https://doi.org/10.1016/j.yjmcc.2007.11.009
Wojakowski W, Kazmierski M, Korzeniowska B (2007) Link between erythropoietin release and mobilization of endothelial progenitor cells in acute myocardial infarction. Eur Heart J 28: 1785-1786. DOI: https://doi.org/10.1093/eurheartj/ehm260
Menasche P, Alfieri O, Janssens S (2008) Autologous Grafting in Ischemic Cardiomyopathy (MAGIC) trial: first randomized placebo-controlled study of myoblast transplantation. Circulation 117: 1189-1200. DOI: https://doi.org/10.1161/CIRCULATIONAHA.107.734103
Dawn B, Guo Y, Rezazadeh A (2006)Postinfarct cytokine therapy regenerates cardiac tissue and improves left ventricular function. Circ Res 98: 1098-1105. DOI: https://doi.org/10.1161/01.RES.0000218454.76784.66
Beltrami AP, Barlucchi L, Torella D (2003)Adult cardiac stem cells are multipotent and support myocardial regeneration. Cell 114: 763-776. DOI: https://doi.org/10.1016/S0092-8674(03)00687-1
Kajstura J, Urbanek K, Rota M (2008)Cardiac stem cells and myocardial disease. J Mol Cell Cardiol;45:505-513. DOI: https://doi.org/10.1016/j.yjmcc.2008.05.025
Bearzi C, Rota M, Hosoda T (2007)Human cardiac stem cells. ProcNatlAcadSci U S A104:14068-14073. DOI: https://doi.org/10.1073/pnas.0706760104
Messina E, De AL, Frati G (2004)Isolation and expansion of adult cardiac stem cells from human and murine heart. Circ Res 95: 911-921. DOI: https://doi.org/10.1161/01.RES.0000147315.71699.51
Smith RR, Barile L, Cho HC (2007) Regenerative potential of cardiosphere-derived cells expanded from percutaneous endomyocardial biopsy specimens. Circulation 115: 896-908. DOI: https://doi.org/10.1161/CIRCULATIONAHA.106.655209
Christensen JL, Weissman IL (2001) Flk-2 is a marker in hematopoietic stem cell differentiation: a simple method to isolate long-term stem cells. ProcNatlAcadSci U S A 98: 14541-14546. DOI: https://doi.org/10.1073/pnas.261562798
Kablar B, Tajbakhsh S, Rudnicki MA (2000)Transdifferentiation of esophageal smooth to skeletal muscle is myogenic bHLH factor-dependent. Development 127: 1627-1639. DOI: https://doi.org/10.1242/dev.127.8.1627
Beltrami AP, Urbanek K, Kajstura J (2001)Evidence that human cardiac myocytes divide after myocardial infarction. N Engl J Med 344: 1750-1757. DOI: https://doi.org/10.1056/NEJM200106073442303
Khaitlina SY (2001) Functional specificity of actin isoforms. Int Rev Cytol 202: 35-98. DOI: https://doi.org/10.1016/S0074-7696(01)02003-4
Curcio F, Ambesi-Impiombato FS, Perrella G (1994)Long-term culture and functional characterization of follicular cells from adult normal human thyroids. ProcNatlAcadSci U S A 91: 9004-9008. DOI: https://doi.org/10.1073/pnas.91.19.9004
Longin A, Souchier C, Ffrench M (1993) Comparison of anti-fading agents used in fluorescence microscopy: image analysis and laser confocal microscopy study. J HistochemCytochem 41: 1833-1840. DOI: https://doi.org/10.1177/41.12.8245431
Donnini D, Perrella G, Stel G (2000) A new model of human aortic endothelial cells in vitro. Biochimie 82:1107-1114. DOI: https://doi.org/10.1016/S0300-9084(00)01195-0
Kimes BW, Brandt BL (1976)Properties of a clonal muscle cell line from rat heart. ExpCellRes 98: 367-381. DOI: https://doi.org/10.1016/0014-4827(76)90447-X
Garlanda C, Parravicini C, Sironi M (1994)Progressive growth in immunodeficient mice and host cell recruitment by mouse endothelial cells transformed by polyoma middle-sized T antigen: implications for the pathogenesis of opportunistic vascular tumors. ProcNatlAcadSci U S A 91: 7291-7295. DOI: https://doi.org/10.1073/pnas.91.15.7291
Buzby JS, Knoppel EM, Cairo MS (1994) Coordinate regulation of Steel factor, its receptor (Kit), and cytoadhesion molecule (ICAM-1 and ELAM-1) mRNA expression in human vascular endothelial cells of differing origins. ExpHematol 22: 122-129.
Kubo H, Jaleel N, Kumarapeli A (2008)Increased cardiac myocyte progenitors in failing human hearts. Circulation 118: 649-657. DOI: https://doi.org/10.1161/CIRCULATIONAHA.107.761031
Matsuda R, Takahashi T, Nakamura S (1993)Expression of the c-kit protein in human solid tumors and in corresponding fetal and adult normal tissues. Am J Pathol 142: 339-346.
De AL, Berghella L, Coletta M (1999)Skeletal myogenic progenitors originating from embryonic dorsal aorta coexpress endothelial and myogenic markers and contribute to postnatal muscle growth and regeneration. J Cell Biol 147: 869-878. DOI: https://doi.org/10.1083/jcb.147.4.869
Quaini F, Urbanek K, Beltrami AP (2002)Chimerism of the transplanted heart. N Engl J Med 346: 5-15. DOI: https://doi.org/10.1056/NEJMoa012081
Torella D, Rota M, Nurzynska D (2004)Cardiac stem cells and myocyte aging, heart failure and insuline-like growth factor-1overexpression. Circ Res; 94:514-524. DOI: https://doi.org/10.1161/01.RES.0000117306.10142.50
Barile L, Messina E, Giacomello A (2007) Endogenous cardiac stem cells. ProgCardiovasc Dis 50: 31-48. DOI: https://doi.org/10.1016/j.pcad.2007.03.005
Niu Z, Iyer D, Conway SJ (2008) Serum response factor orchestrates nascent sarcomerogenesis and silences the biomineralization gene program in the heart. ProcNatlAcadSci U S A 105: 17824-17829. DOI: https://doi.org/10.1073/pnas.0805491105
Tavi P, Pikkarainen S, Ronkainen J (2004)Pacing-induced calcineurin activation controls cardiac Ca2+ signalling and gene expression. J Physiol 554: 309-320. DOI: https://doi.org/10.1113/jphysiol.2003.053579
Sontia B, Touyz RM (2007) Magnesium transport in hypertension. Pathophysiology 14: 205-211. DOI: https://doi.org/10.1016/j.pathophys.2007.09.005
Simpson DG, Decker ML, Clark WA (1993)Contractile activity and cell-cell contact regulate myofibrillar organization in cultured cardiac myocytes. J Cell Biol 123: 323-336. DOI: https://doi.org/10.1083/jcb.123.2.323
Eppenberger-Eberhardt M, Flamme I, Kurer V (1990)Reexpression of alpha-smooth muscle actin isoform in cultured adult rat cardiomyocytes. DevBiol 139: 269-278. DOI: https://doi.org/10.1016/0012-1606(90)90296-U
Kubo H, Berretta RM, Jaleel N (2009)c-Kit+ bone marrow stem cells differentiate into functional cardiac myocytes. Clin Trans Sci2:26-32. DOI: https://doi.org/10.1111/j.1752-8062.2008.00089.x