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. 2023 Jan 10:9:1095141.
doi: 10.3389/fcvm.2022.1095141. eCollection 2022.

Approaches for the isolation and long-term expansion of pericytes from human and animal tissues

Affiliations

Approaches for the isolation and long-term expansion of pericytes from human and animal tissues

Valeria Vincenza Alvino et al. Front Cardiovasc Med. .

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.

Keywords: cardiovascular; isolation and expansion; methods; pericytes; regenerative medicine.

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Conflict of interest statement

The authors declare that the research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Characteristics of pericytes. (A) Immunofluorescence microscopy image of NG2 positive (green) pericytes, with endothelial cells labeled in red and nuclei in blue. (B) Cartoon of pericyte around a capillary lined internally by endothelial cells. (C) Aspect of pericytes in culture. (D) Overlapping of antigens with other mesenchymal cells. For clarity, refer to text for the overlapping with perivascular fibroblasts. (E) Key pericyte functions. Image in (A) has been reproduced with permission from the original article in Avolio et al. (3).
FIGURE 2
FIGURE 2
Advantages and disadvantages of different methods of isolation and purification methods. Figure created in Biorender.com source.

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References

    1. Attwell D, Mishra A, Hall CN, O’Farrell FM, Dalkara T. What is a pericyte? J Cereb Blood Flow Metab. (2016) 36:451–5. 10.1177/0271678X15610340 - DOI - PMC - PubMed
    1. Zimmermann KW. Der feinere bau der blutcapillaren. Z Anat Entwicklungsgeschichte. (1923) 68:29–109. 10.1007/BF02593544 - DOI
    1. Avolio E, Madeddu P. Discovering cardiac pericyte biology: from physiopathological mechanisms to potential therapeutic applications in ischemic heart disease. Vascul Pharmacol. (2016) 86:53–63. 10.1016/j.vph.2016.05.009 - DOI - PubMed
    1. Meijer EM, van Dijk CGM, Kramann R, Verhaar MC, Cheng C. Implementation of pericytes in vascular regeneration strategies. Tissue Eng Part B Rev. (2022) 28:1–21. 10.1089/ten.TEB.2020.0229 - DOI - PubMed
    1. Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. (1973) 52:2745–56. 10.1172/JCI107470 - DOI - PMC - PubMed