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Updated: Aug 12, 2025
Isolation and Purification of Murine Cardiac Pericytes
Published on: August 16, 2019
Approaches for the isolation and long-term expansion of pericytes from human and animal tissues
Valeria Vincenza Alvino1, Khaled Abdelsattar Kassem Mohammed1,2, Yue Gu1
1Bristol Heart Institute, University of Bristol, Bristol, United Kingdom.
Insights
Pericytes are crucial for vascular function and regenerative medicine. Standardizing methods for isolating and defining these cells is essential for developing clinical-grade pericyte therapies.
Area of Science:
- Vascular Biology
- Regenerative Medicine
- Cell Biology
Background:
- Pericytes are perivascular cells integral to capillary and vasa vasorum function.
- Growing clinical interest highlights their roles in vascular control and therapeutic potential.
- Current methods for pericyte isolation and definition lack standardization, hindering reproducibility.
Purpose of the Study:
- To review and critically assess existing methodologies for pericyte definition, isolation, and expansion.
- To propose improvements for standardization and harmonization of pericyte research and manufacturing.
- To emphasize the need for clinical-grade pericyte products compliant with regulatory guidelines.
Main Methods:
- Review of current literature on pericyte isolation, characterization, and expansion techniques.
- Analysis of discrepancies and challenges in defining bona fide pericytes.
- Discussion of integrated approaches including RNA-sequencing and single-cell spatial analysis.
Main Results:
- Significant variability exists in methods, impacting reproducibility and the development of pure pericyte preparations.
- Challenges in defining pericytes due to their phenotypic plasticity and heterogeneity.
- Limited progress in developing pericytes as clinical-grade therapeutic products.
Conclusions:
- Standardization of pericyte isolation and characterization protocols is critical for advancing clinical applications.
- Development of robust manufacturing processes is necessary for therapeutic pericyte products.
- Integrating advanced techniques like single-cell spatial analysis and RNA-seq can unlock pericyte potential in health and disease.
Abstract:
Pericytes surround capillaries in every organ of the human body. They are also present around the vasa vasorum, the small blood vessels that supply the walls of larger arteries and veins. The clinical interest in pericytes is rapidly growing, with the recognition of their crucial roles in controlling vascular function and possible therapeutic applications in regenerative medicine. Nonetheless, discrepancies in methods used to define, isolate, and expand pericytes are common and may affect reproducibility. Separating pure pericyte preparations from the continuum of perivascular mesenchymal cells is challenging. Moreover, variations in functional behavior and antigenic phenotype in response to environmental stimuli make it difficult to formulate an unequivocal definition of bona fide pericytes. Very few attempts were made to develop pericytes as a clinical-grade product. Therefore, this review is devoted to appraising current methodologies' pros and cons and proposing standardization and harmonization improvements. We highlight the importance of developing upgraded protocols to create therapeutic pericyte products according to the regulatory guidelines for clinical manufacturing. Finally, we describe how integrating RNA-seq techniques with single-cell spatial analysis, and functional assays may help realize the full potential of pericytes in health, disease, and tissue repair.
