Highly contractile 3D tissue engineered skeletal muscles from human iPSCs reveal similarities with primary myoblast-derived tissues.

Skeletal muscle research is transitioning toward 3D tissue engineered in vitro models reproducing muscle's native architecture and supporting measurement of functionality. Human induced pluripotent stem cells (hiPSCs) offer high yields of cells for differentiation. It has been difficult to differentiate high-quality, pure 3D muscle tissues from hiPSCs that show contractile properties comparable to primary myoblast-derived tissues. Here, we present a transgene-free method for the generation of purified, expandable myogenic progenitors (MPs) from hiPSCs grown under feeder-free conditions. We defined a protocol with optimal hydrogel and medium conditions that allowed production of highly contractile 3D tissue engineered skeletal muscles with forces similar to primary myoblast-derived tissues. Gene expression and proteomic analysis between hiPSC-derived and primary myoblast-derived 3D tissues revealed a similar expression profile of proteins involved in myogenic differentiation and sarcomere function. The protocol should be generally applicable for the study of personalized human skeletal muscle tissue in health and disease.

Efficacy and safety of anterior vaginal prolapse treatment using single incision repair system: Multicentric study.

OBJECTIVE: Up to 80% of all POP surgical procedures are due to anterior vaginal wall prolapse. The aim of this study is to evaluate the efficacy and safety of transvaginal anterior mesh for POP surgical repair. MATERIALS AND METHODS: 153 consecutive patients with symptomatic or recurrent anterior vaginal prolapse undergoing surgical single-incision mesh (Calistar S) were prospectively enrolled in the study. Preoperative evaluation was performed collecting urogynecological history and performing a clinical exam, 3-day voiding diary and urodynamic testing. All incontinent patients completed the International Consultation on Incontinence Questionnaire - Short Form (ICIQ-SF). Operative time, blood loss, perioperative and postoperative complications were prospectively recorded. Postoperative follow-up was scheduled at 1, 6, and 12 months with a urogynecological interview and examination. Success rate was assessed at 1, 6 and 12 months postoperatively. RESULTS: The median follow-up was 16.4 months. None of patients had intraoperative complications. Eight patients (5%) required surgical intervention for complications (5 patients (3%) for haematoma and 3 (2%) for vaginal erosion). At 12 months of follow up 130 out of 140 patients (93%) gained the subjective cure criterion, while 129 out of 140 patients (92%) obtained the objective cure criterion. Eleven (7.8%) patients experienced stage 2 or higher prolapse recurrence and three of them with a stage ≥3 underwent reintervention. No significant differences were recorded in primary outcome at 1, 6 or 12 months postoperatively. CONCLUSIONS: Anterior compartment prolapse repair by Calistar S (single-incision vaginal mesh) is an effective and safe procedure without significant complications.

Novel GAA Variants and Mosaicism in Pompe Disease Identified by Extended Analyses of Patients with an Incomplete DNA Diagnosis.

Pompe disease is a metabolic disorder caused by a deficiency of the glycogen-hydrolyzing lysosomal enzyme acid α-glucosidase (GAA), which leads to progressive muscle wasting. This autosomal-recessive disorder is the result of disease-associated variants located in the GAA gene. In the present study, we performed extended molecular diagnostic analysis to identify novel disease-associated variants in six suspected Pompe patients from four different families for which conventional diagnostic assays were insufficient. Additional assays, such as a generic-splicing assay, minigene analysis, SNP array analysis, and targeted Sanger sequencing, allowed the identification of an exonic deletion, a promoter deletion, and a novel splicing variant located in the 5' UTR. Furthermore, we describe the diagnostic process for an infantile patient with an atypical phenotype, consisting of left ventricular hypertrophy but no signs of muscle weakness or motor problems. This led to the identification of a genetic mosaicism for a very severe GAA variant caused by a segmental uniparental isodisomy (UPD). With this study, we aim to emphasize the need for additional analyses to detect new disease-associated GAA variants and non-Mendelian genotypes in Pompe disease where conventional DNA diagnostic assays are insufficient.