Abstract
Human muscle biopsies are increasingly important for diagnosis, research, and to monitor therapeutic trials. We examined the use of a self-contained, vacuum-assisted biopsy system and a novel muscle freezing technique to improve, simplify, and standardize human muscle biopsy collection and cryopreservation in older adults.The VACORA vacuum-assisted biopsy system was deployed in muscle biopsies of 12 individuals ranging in age from 57 to 80 years. This office-based approach was well tolerated as it is minimally invasive, uses only local anesthetic, and has a quick recovery. To maximize biopsy sample quality and reproducibility, we developed a novel muscle sample freezing protocol. Fresh muscle biopsy samples were placed into readily available tissue cassettes followed by direct freezing in liquid nitrogen. After this modified snap freezing protocol, frozen muscle samples were enrobed in embedding medium for cryosectioning. We examined the effect of this freezing approach in histological sections of rodent and human muscle samples.The VACORA Biopsy System provided as many as four skeletal muscle core samples from a single biopsy site. Biopsy samples from 12 older adults weighed an average of 147.5 ± 11 mg each and had a consistent size and shape. There were no complications, and the residual scar is less than 10 mm. The freezing method using standard tissue cassettes with direct freezing in liquid nitrogen yielded high quality cryopreserved muscle tissue suitable for histological analysis without the need for isopentane and with little to no freeze-thaw damage.These enhancements have streamlined and improved the consistency of our muscle biopsy protocol and provide sufficient high-quality sample for multi-dimensional downstream studies of human muscle in aging and disease.
Highlights
The worldwide prevalence of neuromuscular disease is growing and approximates that of other diseases such as Parkinson’s [1]
Basic research, and therapeutic trials will benefit from ready access to human skeletal muscle samples from a wide range of subjects
Human muscle biopsies were obtained with informed consent from patients of the UCLA Center for Duchenne Muscular Dystrophy (CDMD) under University of California Los Angeles IRB‐approved protocols #11‐001087 and #18-001547
Summary
The worldwide prevalence of neuromuscular disease is growing and approximates that of other diseases such as Parkinson’s [1]. Basic research, and therapeutic trials will benefit from ready access to human skeletal muscle samples from a wide range of subjects. Diagnosis of neuromuscular disorders often relies on clinical observations, but is accelerated with molecular studies of the muscle itself. Older adults may have muscle biopsies to investigate acute muscle weakness, but the unrelenting age-induced declines in muscle mass or function rarely prompt a muscle biopsy. This is despite our current lack of a clear etiology for muscle aging. The advent of therapeutic interventions for some neuromuscular diseases requires validated biomarkers of treatment efficacy
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.