从肌肉活检患者来源的细胞系疾病建模隔离和永生化

Overview

This protocol describes techniques for live cell isolation and primary culture of myogenic and fibroblast cell lines from muscle or skin tissue. The overall goal is to isolate purified muscle cells from human patient muscle biopsies and immortalize them for downstream studies.

Key Study Components

Area of Science

• Cell Biology
• Muscle Physiology
• Regenerative Medicine

Background

• Isolation of patient-derived cells is crucial for studying muscle diseases.
• Immortalized cell lines can provide unlimited cells for research.
• Techniques for cell purification and culture are essential for downstream applications.
• Myogenic cells can be differentiated and expanded for various studies.

Purpose of Study

• To isolate and immortalize muscle cells from human biopsies.
• To provide a reliable tool for generating patient-derived cells.
• To enable high-throughput drug screening and research applications.

Methods Used

• Weighing, mincing, and digesting tissue to dissociate cells.
• Purification of myogenic cells using FACS.
• Infection of myogenic cells with CDK4 and Hturt for immortalization.
• Assessment of cell fusion and expression of myogenic markers.

Main Results

• Patient-derived muscle cells can be expanded for several passages.
• Cells can differentiate based on cell fusion assays.
• Presence of twitching myotubes indicates successful differentiation.
• Expression of myogenic markers confirms cell identity.

Conclusions

• This technique provides a reliable source of myogenic cells.
• It has implications for muscle disease diagnosis and treatment.
• Potential for unlimited cell supply for various research applications.

Frequently Asked Questions