Isolation and characterization of myogenic precursor cells from human cremaster muscle

Human myogenic precursor cells have been isolated and expanded from a number of skeletal muscles, but alternative donor biopsy sites must be sought after in diseases where muscle damage is widespread. Biopsy sites must be relatively accessible, and the biopsied muscle dispensable. Here, we aimed to histologically characterize the cremaster muscle with regard number of satellite cells and regenerative fibres, and to isolate and characterize human cremaster muscle-derived stem/precursor cells in adult male donors with the objective of characterizing this muscle as a novel source of myogenic precursor cells. Cremaster muscle biopsies (or adjacent non-muscle tissue for negative controls; N=19) were taken from male patients undergoing routine surgery for urogenital pathology. Myosphere cultures were derived and tested for their in vitro and in vivo myogenic differentiation and muscle regeneration capacities. Cremaster-derived myogenic precursor cells were maintained by myosphere culture and efficiently differentiated to myotubes in adhesion culture. Upon transplantation to an immunocompromised mouse model of cardiotoxin-induced acute muscle damage, human cremaster-derived myogenic precursor cells survived to the transplants and contributed to muscle regeneration. These precursors are a good candidate for cell therapy approaches of skeletal muscle. Due to their location and developmental origin, we propose that they might be best suited for regeneration of the rhabdosphincter in patients undergoing stress urinary incontinence after radical prostatectomy.We thank patients and medical personnel for their generous involvement in the study. We also acknowledge the help of Biodonostia Animal and Experimental Operations Facility. This work was supported by grants from Ministerio de Economia y Competitividad (RTC-2015-3750-1) and Instituto de Salud Carlos III (PI13/02172, PI16/01430) to A.I., co-funded by the European Union (ERDF/ESF, 'Investing in your future'). N.N.-G. received a studentship from the Department of Education, University and Research of the Basque Government (PRE2013-1-1168). A.L.M. was funded by grants from FIS (PI17/01841 and PI14/00436), CIBERNED and the Basque Government (2015/11038, RIS3 2017222021 and BIO16/ER/022). M.F.L.-C. was supported by the Servicio Andaluz de Salud from the Consejeria de Salud de la Junta de Andalucia, grant PI 0222-2014, co-funded by the European Union (ERDF/ESF). I.M.A was funded by grants from Ministerio de Economia y Competitividad (PEJ-2014-P-01215 and FJCI-2016-28121)

Patient-Specific iPSC-Derived Cellular Models of LGMDR1

Limb-girdle muscular dystrophy recessive 1 (LGMDR1) represents one of the most common types of LGMD in the population, where patients develop a progressive muscle degeneration. The disease is caused by mutations in calpain 3 gene, with over 500 mutations reported to date. However, the molecular events that lead to muscle wasting are not clear, nor the reasons for the great clinical variability among patients, and this has so far hindered the development of effective therapies. Here we generate human induced pluripotent stem cells (iPSCs) from skin fibroblasts of 2 healthy controls and 4 LGMDR1 patients with different mutations. The generated lines were able to differentiate into myogenic progenitors and myotubes in vitro and in vivo, upon a transient PAX7 overexpressing protocol. Thus, we have generated myogenic cellular models of LGMDR1 that harbor different CAPN3 mutations within a human genetic background, and which do not derive from muscular biopsies. These models will allow us to investigate disease mechanisms and test therapies. Despite the variability found among iPSC lines that was unrelated to CAPN3 mutations, we found that patient-derived myogenic progenitors and myotubes express lower levels of DMD, which codes a key protein in satellite cell regulation and myotube maturation.This work has been funded by grants from Ilundain Foundation, Isabel Gemio Foundation, Fundació La Caixa, Basque Government (2015111038), Catalan Government (2017-SGR-899 and CERCA Programme), Provincial Council of Gipuzkoa (A.LdM 114/17), and Instituto de Salud Carlos III (PI14/00436, PS09/00660 and RD16/0011/0024). A.M.-A and N.N.-G. received a studentship from the Department of Education, University and Research of the Basque Government (BFI-2012-19, PRE2013-1-1168

[Montreal 1976] [Material gráfico]

Contiene fotografías pertenecientes al archivo fotográfico del diario "Región", publicadas entre 1974 y 1976, aunque la mayoría en 1976Todas las fotografías firmadas por Foto E. Gar (Oviedo), Cifra Gráfica, y EF