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Cell Biology JoVE (Open Access) Citable · DOI

Purification of Progenitors from Skeletal Muscle

DOI: 10.3791/2476-v
What you'll learn
  • ✓ Dissociate skeletal muscle into single cells
  • ✓ Label fibro/adipogenic and myogenic progenitors
  • ✓ Purify progenitors by flow cytometry
  • ✓ Transplant sorted progenitors into recipients
Protocol

Biopharma Insights Method for the enzymatic dissociation, surface labeling and purification by flow cytometry of fibro/adipogenic and myogenic progenitors from murine skeletal muscle.

Difficulty
advanced
Total time
~6-8 hrs per muscle sample
Model organism
Mouse
Biosafety
BSL-1

Steps

1
Collect skeletal muscle tissue

Harvest muscle tissue from donor mice, ensuring sterile conditions and appropriate animal handling.

▶ 02:35
2
Enzymatically digest muscle into single cells

Incubate minced muscle with digestive enzymes to release individual cells, then filter to obtain a single-cell suspension.

▶ 03:49
3
Stain cells with antibodies and sort

Label cells with fluorophore-conjugated antibodies against progenitor markers, then use flow cytometry to separate fibro/adipogenic and myogenic progenitor populations.

▶ 05:52
4
Transplant sorted progenitor cells

Inject purified progenitor cells into recipient mice for functional analysis of their regenerative potential.

▶ 08:45
5
Analyze isolation and transplantation results

Evaluate the purity and functionality of sorted progenitors by examining representative myogenic and adipogenic outcomes post-transplantation.

▶ 09:45
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