The Duchenne and Becker muscular dystrophies are caused by mutation of dystrophin gene and primar... more The Duchenne and Becker muscular dystrophies are caused by mutation of dystrophin gene and primarily affect skeletal and cardiac muscles. Cardiac involvement in dystrophic GRMD dogs has been demonstrated by electrocardiographic studies with the onset of a progressive cardiomyopathy similar to the cardiac disease in DMD patients. In this respect, GRMD is a useful model to explore cardiac and skeletal muscle pathogenesis and for developing new therapeutic protocols. Here we describe a protocol to convert GRMD canine fibroblasts isolated from heart and skin into induced cardiac-like myocytes (ciCLMs). We used a mix of transcription factors (GATA4, HAND2, TBX5, and MEF2C), known to be able to differentiate mouse and human somatic cells into ciCLMs. Exogenous gene expression was obtained using four lentiviral vectors carrying transcription factor genes and different resistance genes. Our data demonstrate a direct switch from fibroblast into ciCLMs with no activation of early cardiac gene...
Mesoangioblasts (MABs) are mesoderm-derived stem cells, associated with small vessels and origina... more Mesoangioblasts (MABs) are mesoderm-derived stem cells, associated with small vessels and originally described in the mouse embryonic dorsal aorta. Similar though not identical cells have been later identified and characterized from postnatal small vessels of skeletal muscle and heart. They have in common the expression of pericyte markers, the anatomical location, the ability to self-renew in culture, and to differentiate into various types of mesodermal lineages upon proper culture conditions. Currently, the developmental origin of MABs and the relationship with other muscle stem cells are not understood in detail and are the subject of active research. This chapter provides an outline of the latest techniques for isolation and characterization of adult MABs from human and mouse skeletal muscles.
Regenerative medicine for skeletal and cardiac muscles still constitutes a fascinating and ambiti... more Regenerative medicine for skeletal and cardiac muscles still constitutes a fascinating and ambitious frontier. In this perspective, understanding the possibilities of intrinsic cell plasticity, present in post-natal muscles, is vital to define and improve novel therapeutic strategies for acute and chronic diseases. In addition, many somatic stem cells are now crossing the boundaries of basic/translational research to enter the first clinical trials. However, it is still an open question whether a lineage switch between skeletal and cardiac adult myogenesis is possible. Therefore, this review focuses on resident somatic stem cells of post-natal skeletal and cardiac muscles and their plastic potential toward the two lineages. Furthermore, examples of myogenic lineage switch in adult stem cells are also reported and discussed.
The Duchenne and Becker muscular dystrophies are caused by mutation of dystrophin gene and primar... more The Duchenne and Becker muscular dystrophies are caused by mutation of dystrophin gene and primarily affect skeletal and cardiac muscles. Cardiac involvement in dystrophic GRMD dogs has been demonstrated by electrocardiographic studies with the onset of a progressive cardiomyopathy similar to the cardiac disease in DMD patients. In this respect, GRMD is a useful model to explore cardiac and skeletal muscle pathogenesis and for developing new therapeutic protocols. Here we describe a protocol to convert GRMD canine fibroblasts isolated from heart and skin into induced cardiac-like myocytes (ciCLMs). We used a mix of transcription factors (GATA4, HAND2, TBX5, and MEF2C), known to be able to differentiate mouse and human somatic cells into ciCLMs. Exogenous gene expression was obtained using four lentiviral vectors carrying transcription factor genes and different resistance genes. Our data demonstrate a direct switch from fibroblast into ciCLMs with no activation of early cardiac gene...
Mesoangioblasts (MABs) are mesoderm-derived stem cells, associated with small vessels and origina... more Mesoangioblasts (MABs) are mesoderm-derived stem cells, associated with small vessels and originally described in the mouse embryonic dorsal aorta. Similar though not identical cells have been later identified and characterized from postnatal small vessels of skeletal muscle and heart. They have in common the expression of pericyte markers, the anatomical location, the ability to self-renew in culture, and to differentiate into various types of mesodermal lineages upon proper culture conditions. Currently, the developmental origin of MABs and the relationship with other muscle stem cells are not understood in detail and are the subject of active research. This chapter provides an outline of the latest techniques for isolation and characterization of adult MABs from human and mouse skeletal muscles.
Regenerative medicine for skeletal and cardiac muscles still constitutes a fascinating and ambiti... more Regenerative medicine for skeletal and cardiac muscles still constitutes a fascinating and ambitious frontier. In this perspective, understanding the possibilities of intrinsic cell plasticity, present in post-natal muscles, is vital to define and improve novel therapeutic strategies for acute and chronic diseases. In addition, many somatic stem cells are now crossing the boundaries of basic/translational research to enter the first clinical trials. However, it is still an open question whether a lineage switch between skeletal and cardiac adult myogenesis is possible. Therefore, this review focuses on resident somatic stem cells of post-natal skeletal and cardiac muscles and their plastic potential toward the two lineages. Furthermore, examples of myogenic lineage switch in adult stem cells are also reported and discussed.
Uploads
Papers