Human Skeletal Muscle Cells (DMD)
Cat.No.: CSC-C3604
Species: Human
Source: Skeletal Muscle
Cell Type: Skeletal Muscle Cell
- Specification
- Background
- Scientific Data
- Q & A
- Customer Review
DMD is a recessive X-linked form of muscular dystrophy characterized by rapid progression of muscle degeneration.
Origin: Human
Disease: Duchenne Muscular Dystrophy (DMD)
Application: These cells can be used for various types of in vitro, in vivo, or regenerative medicine studies in normal or diseased systems and in muscle development studies.
Note: Never can cells be kept at -20 °C.
Duchenne muscular dystrophy (DMD) is a fatal X-linked recessive disease that affects 1 in 5.000 newborn males. It is typically caused by deletions disrupting the open reading frame of the DMD gene, resulting in a lack of dystrophin protein. Dystrophin plays a major role in membrane stabilization during muscle contraction, linking the actin cytoskeleton to the sarcolemma and also contributes to extracellular signalling. The lack of dystrophin in the muscles of DMD patients leads to progressive muscle wasting and degeneration. Currently, there is no curative treatment for DMD.
In vitro cell models are particularly useful for evaluating the efficiency of novel therapies for DMD. However, only a few human immortalized muscle cell lines derived from DMD patients are currently available. Due to the wide spectrum of DMD mutations and the difficulties to obtain DMD patient muscle biopsies, DMD-myoblasts models would provide a powerful resource for in vitro drug screening and study disease rescue mechanisms.
Creative Bioarray offers single-donor Primary Human Skeletal Muscle Cells and Skeletal Muscle Myoblasts, characterized by age, gender, race, and BMI with specially formulated media that is optimized to make cell culture growth, differentiation, and cryopreservation as reliable as possible in your lab.
Amelioration of Morphological Pathology in Cardiac, Respiratory, and Skeletal Muscles Following Intraosseous Administration of Human Dystrophin Expressing Chimeric (DEC) Cells in Duchenne Muscular Dystrophy Model
Background
This study evaluates human Dystrophin Expressing Chimeric (DEC) cell therapy administered via systemic-intraosseous route in mdx/scid mice, a Duchenne muscular dystrophy model. Histological and morphometric analyses of cardiac, diaphragm, and gastrocnemius muscles were performed at 90 and 180 days after different DEC doses. Using Hematoxylin & Eosin staining and minimal Feret’s diameter measurement, DEC-treated mice showed improved muscle morphology, with fiber size distribution shifting toward wild-type phenotype and increased mean fiber diameters compared to vehicle controls. Results confirm long-term efficacy and dose-dependent normalization of dystrophic pathology across critical muscles.
Key Findings
The study assessed human DEC cell therapy delivered systemically-intraosseously in mdx/scid mice at 90 and 180 days. In cardiac muscle, DEC treatment significantly reduced fibers below 20 μm and increased fibers within 20–50 μm, with mean Feret's diameters rising to wild-type levels (e.g., 16.76 μm at 90 days for 1×10^6 cells vs. 14.64 μm vehicle). Diaphragm showed increased mid-sized fibers and reduced small/large fibers, with higher dose approaching wild-type. In gastrocnemius, DEC shifted fiber distribution toward larger sizes and significantly increased mean diameter at 5×10^6 cells at 180 days (34.01 μm vs. 28.40 μm vehicle). Overall, DEC therapy ameliorated dystrophic pathology across cardiac, respiratory, and skeletal muscles in a dose-dependent manner, confirming long-term morphological efficacy.
Contribution
Creative Bioarray supplied DMD-affected human myoblasts (cat. CSC-C3604) that were fused with normal human myoblasts to generate dystrophin-expressing chimeric (DEC) cells. This critical cell source enabled the preclinical evaluation of systemic-intraosseous DEC therapy in the mdx/scid Duchenne muscular dystrophy model, supporting the study's morphological and therapeutic findings.
Circadian Clock Control of Muscle Stem Cells Through Temporal Coordination of Notch and Wnt Signaling
Background
The circadian clock regulates stem cell responses during tissue remodeling. In skeletal muscle regeneration, a temporally coordinated transition from Notch to Wnt signaling is required for proper myogenesis, but the mechanisms linking clock to this switch were unknown. This study shows that circadian clock activity marks cycling myogenic progenitors and directly controls Notch pathway components. Clock coordinates Notch and Wnt signaling to drive myogenic progression. Genetic and pharmacological clock activation enhanced progenitor proliferation and differentiation, and improved regeneration in dystrophic muscle. These findings establish the clock as a temporal regulator of regenerative signaling, offering potential targeted interventions.
Key Findings
Circadian clock activity marks the cycling population of regeneration-activated myogenic progenitors, induced in concert with Notch signaling. Key Notch pathway components (Notch1, Jag1, HeyL) are direct Bmal1 transcriptional targets via E-box binding. The clock coordinates Notch and Wnt signaling to drive myogenic progression. Genetic activation of clock in satellite cells (Bmal1 overexpression) and pharmacological clock stimulation (chlorhexidine and CM002) enhanced proliferative expansion and subsequent differentiation of myogenic progenitors during regeneration. Effects were mediated by early Notch activation followed by increased Wnt activity at later stages. Both clock-dependent mechanisms remained functional in dystrophin-deficient mouse muscle and human myoblasts. Clock-activating compounds enhanced regenerative myogenesis after acute injury and improved regeneration in dystrophic muscle. These findings establish the circadian clock as a temporal regulator of regenerative signaling programs.
Contribution
Creative Bioarray provided human DMD primary myoblasts (CSC-C3604) used to test clock-activating compounds on myogenic differentiation. These cells enabled validation of clock-dependent mechanisms in a human dystrophic disease context, demonstrating that pharmacological clock activation promotes myogenesis in DMD myoblasts.
Characterizing Primary Donor Cells from Duchenne Muscular Dystrophy (DMD) Patients
The process of donor-specific myoblast selection and cell expansion is illustrated in Fig. 1A. In all, cells from 4 donors were selected with 2 coming from DMD patients and the other 2 coming from non-DMD individuals.
For further characterization, the cells were differentiated and stained for Myosin heavy chain (MHC) to identify myotubes (Fig. 1B). Cell images were then used to extract the myotube area (MHC-positive area), and nuclei count (Fig. 1C), which was used to compute the fusion index. As expected, the fusion index for the DMD donors (DMD #1 and DMD #4) was significantly lower compared to non-DMD donors (Non-DMD #1 and Non-DMD #3).
Dystrophin expression was characterized by immunofluorescence (IF) and simple western blot (Fig. 1D-1F). Nuclei and myotubes were identified using Hoechst and Troponin T respectively. The DMD donors have very low to no expression of Dystrophin compared to the non-DMD donors.
Fig. 3. Donor selection, myotube quantification and dystrophin expression validation (Hariharan, Santosh, et al. 2023).
Intracellular CD56 Intensity Per Nuclei Does Not Correlate with The Fusion Index
Skeletal muscle-derived cells (SMDC) hold tremendous potential for replenishing dysfunctional muscle lost due to disease or trauma. Predicting the therapeutic potency of SMDC in vitro prior to implantation is key to developing successful therapeutics in regenerative medicine and reducing implementation costs. The primary aim of the study was thus precisely to evaluate the applicability of multi-parametric imaging-based phenotypic characterisation to distinguish the myogenic potency of SMDC obtained from different donors.
CD56 is a reliable marker for myoblasts among cultured cells from skeletal muscle and it was reasoned that its level of expression could reflect the fusion index in cells derived from different donors. However, the mean intracellular CD56 intensity per nuclei did not correlate with the fusion index after 5 days of differentiation and the range of CD56 intracellular staining intensity between donors was small (Fig. 2).
Fig. 4. CD56 intracellular expression (Desprez, Charlotte, et al. 2023).
Population doubling is 80-120 or up to 12 passages when cultured at 60% to 70% confluent in Creative Bioarray culture system.
Ask a Question
Write your own review
- You May Also Need
Description: HSkMSC from Creative Bioarray are isolated from human muscle of the pectoral girdle. HSkMSC are cryopreserved at primary culture or passage one and delivered frozen. Each vial contains >5 x 10^5 ...
Description: Origin: HumanDesease: Duchenne Muscular Dystrophy (DMD)Application: These cells can be used for various types of in vitro, in vivo, or regenerative medicine studies in normal or diseased systems and ...
Description: Human skeletal muscle myoblasts are derived from whole muscle that has been dissociated into single cells and purified using differential adhesion. Human skeletal muscle myoblasts are from a single ...
Description: Human Skeletal Muscle Microvascular Endothelial Cells are isolated from human skeletal muscle tissue.
Description: Diabetic Type I and Type II Human Skeletal Muscle Myoblasts (HSMM) are isolated from donors diagnosed with either Diabetes Type I or Diabetes Type II. Additional donor information is available by ...
Description: Diabetic Type I and Type II Human Skeletal Muscle Myoblasts (HSMM) are isolated from donors diagnosed with either Diabetes Type I or Diabetes Type II. Additional donor information is available by ...