On May 5, 2024, the Bio-protocol Journal published online a method article entitled "Generation of Skeletal Muscle Organoids from Human Pluripotent Stem Cells" by the Holm Zaehres team of Ruhr-Universit ä t Bochum in Bochum, Germany.

Key words: skeletal muscle organ myogenesis induces pluripotent stem cell satellite cells PAX7 disease model muscular dystrophy

Organ-like (Organoids) refers to tissue analogues with certain spatial structure formed by three-dimensional (3D) culture of adult stem cells or pluripotent stem cells in vitro.Although the quasi-organ is not a real human organ, it can simulate the real organ in structure and function, simulate the tissue structure and function of the body to the greatest extent, and can be cultured stably for a long time.

一. Background introduction

In the study of human disease models, in vitro models based on human stem cells gradually replace the traditional animal models, especially in the field of hereditary diseases. This method uses patient-specific cells for reprogramming, so that the corresponding cell lines can be generated for each genetic disease, and the pathological phenotype and its molecular mechanism can be further studied. The Food and Drug Administration's Modernization Act 2.0 has also promoted the development of this area, encouraging the use of cell culture techniques for the testing of new drugs. The differentiation of pluripotent stem cells (PSCs) has been proved to be effective in guiding these cells to differentiate into skeletal muscle cells, which involves activating WNT pathway, inhibiting BMP pathway and applying FGF and other factors. Recently, three-dimensional (3D) differentiation technology has become a research hotspot because it can simulate the complex interaction and microenvironment between cells, which is of great significance for understanding and simulating the pathogenesis of muscular degenerative diseases such as Duchenne muscular dystrophy.

二. Highlight of the method. 

1. This method can be used to analyze muscle formation from early embryo to fetal stage, which provides a powerful tool for studying muscle development. 

2. This system can provide skeletal muscle progenitor cells in vitro, and these cells can be maintained in the incubator for up to 14 weeks, which is of great significance for the study of long-term muscle development and pathological changes. 

3. The use of patient-specific induced pluripotent stem cells (iPSCs) can bypass the bottleneck of muscle biopsy in patients and provide a new way to study human myogenesis in healthy and diseased states.

三. Potential application. 

1. Disease modeling and research. 

Skeletal muscle organs produced by patients' specific induced pluripotent stem cells (iPSCs) can simulate the development of various hereditary muscle diseases, such as Duchenne muscular dystrophy, to study the pathological mechanism and early phenotype. 

2. Drug testing and screening. 

By simulating the growth and pathological changes of human muscle in vitro, this system can be used to test and screen potential therapeutic drugs and speed up the drug development process. 

3. Gene therapy research. 

Using this model, we can explore and optimize gene therapy strategies for specific genetic diseases, such as editing specific genes through CRISPR/Cas9 technology.

四. Repeatability verification. 

The repeatability of this method has been verified by the Holm Zaehres team of Ruhr-Universit ä t Bochum, Germany, and the relevant experimental data and results are published in Elife,DOI:10.1093/nar/gku1255.