Adult cardiac stem cells are multipotent and robustly myogenic: c-kit expression is necessary but not sufficient for their identification.

Multipotent adult resident cardiac stem cells (CSCs) were first identified by the expression of c-kit, the stem cell factor receptor. However, in the adult myocardium c-kit alone cannot distinguish CSCs from other c-kit-expressing (c-kitpos) cells. The adult heart indeed contains a heterogeneous mixture of c-kitpos cells, mainly composed of mast and endothelial/progenitor cells. This heterogeneity of cardiac c-kitpos cells has generated confusion and controversy about the existence and role of CSCs in the adult heart. Here, to unravel CSC identity within the heterogeneous c-kit-expressing cardiac cell population, c-kitpos cardiac cells were separated through CD45-positive or -negative sorting followed by c-kitpos sorting. The blood/endothelial lineage-committed (Lineagepos) CD45posc-kitpos cardiac cells were compared to CD45neg(Lineageneg/Linneg) c-kitpos cardiac cells for stemness and myogenic properties in vitro and in vivo. The majority (~90%) of the resident c-kitpos cardiac cells are blood/endothelial lineage-committed CD45posCD31posc-kitpos cells. In contrast, the LinnegCD45negc-kitpos cardiac cell cohort, which represents ⩽10% of the total c-kitpos cells, contain all the cardiac cells with the properties of adult multipotent CSCs. These characteristics are absent from the c-kitneg and the blood/endothelial lineage-committed c-kitpos cardiac cells. Single Linnegc-kitpos cell-derived clones, which represent only 1-2% of total c-kitpos myocardial cells, when stimulated with TGF-ß/Wnt molecules, acquire full transcriptome and protein expression, sarcomere organisation, spontaneous contraction and electrophysiological properties of differentiated cardiomyocytes (CMs). Genetically tagged cloned progeny of one Linnegc-kitpos cell when injected into the infarcted myocardium, results in significant regeneration of new CMs, arterioles and capillaries, derived from the injected cells. The CSC's myogenic regenerative capacity is dependent on commitment to the CM lineage through activation of the SMAD2 pathway. Such regeneration was not apparent when blood/endothelial lineage-committed c-kitpos cardiac cells were injected. Thus, among the cardiac c-kitpos cell cohort only a very small fraction has the phenotype and the differentiation/regenerative potential characteristics of true multipotent CSCs.

Methods and Techniques for miRNA Data Analysis.

Genomic data analysis consists of techniques to analyze and extract information from genes. In particular, genome sequencing technologies allow to characterize genomic profiles and identify biomarkers and mutations that can be relevant for diagnosis and designing of clinical therapies. Studies often regard identification of genes related to inherited disorders, but recently mutations and phenotypes are considered both in diseases studies and drug designing as well as for biomarkers identification for early detection. Gene mutations are studied by comparing fold changes in a redundancy version of numeric and string representation of analyzed genes starting from macromolecules. This consists of studying often thousands of repetitions of gene representation and signatures identified by biological available instruments that starting from biological samples generate arrays of data representing nucleotides sequences representing known genes in an often not well-known sequence. High-performance platforms and optimized algorithms are required to manipulate gigabytes of raw data that are generated by the so far mentioned biological instruments, such as NGS (standing for Next-Generation Sequencing) as well as for microarray. Also, data analysis requires the use of several tools and databases that store gene targets as well as gene ontologies and gene-disease association. In this chapter we present an overview of available software platforms for genomic data analysis, as well as available databases with their query engines.