Home › Immunology › Isolation and Characterization of Cardiac Mesenchymal Stromal Cells from Endomyocardial Bioptic Samples of Arrhythmogenic Cardiomyopathy Patients
Immunology JoVE (Open Access) Citable · DOI

Isolation and Characterization of Cardiac Mesenchymal Stromal Cells from Endomyocardial Bioptic Samples of Arrhythmogenic Cardiomyopathy Patients

DOI: 10.3791/57263-v
What you'll learn
  • ✓ Isolate cardiac mesenchymal stromal cells from endomyocardial biopsy samples
  • ✓ Characterize C-MSCs using flow cytometry marker analysis
  • ✓ Induce and verify adipogenic differentiation via Oil Red O staining
Protocol

Biopharma Insights In this article, a method to isolate cardiac mesenchymal stromal cells from endomyocardial bioptic samples of arrhythmogenic cardiomyopathy patients is provided. Their characterization and the protocol to boost their adipogenic differentiation are described.

Difficulty
advanced
Total time
~3–5 days (biopsy collection, cell isolation, culture expansion, differentiation, staining)
Model organism
Human (arrhythmogenic cardiomyopathy patient endomyocardial biopsies)
Biosafety
BSL-2

Steps

1
Collect and process cardiac biopsy tissue

Obtain endomyocardial bioptic samples from arrhythmogenic cardiomyopathy patients and prepare tissue for cell isolation. Enzymatic digestion and mechanical dissociation are performed to release stromal cells.

▶ 01:12
2
Characterize cardiac MSCs by flow cytometry

Perform phenotypic analysis of isolated C-MSCs using fluorescence-labeled antibodies against mesenchymal and hematopoietic markers to confirm cell identity and surface marker expression patterns.

▶ 03:06
3
Stain differentiated cells with Oil Red O

Culture C-MSCs under adipogenic induction conditions and apply Oil Red O staining to visualize and confirm lipid accumulation in differentiated adipocytes.

▶ 05:22
4
Analyze flow cytometry and staining results

Quantify flow cytometry data and assess Oil Red O staining intensity to evaluate mesenchymal marker expression and adipogenic differentiation efficiency.

▶ 07:02
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