Mitigating the non-specific uptake of immunomagnetic microparticles enables the extraction of endothelium from human fat

Jeremy A Antonyshyn,Stefan O P Hofer,Craig A Simmons,Meghan J Mcfadden,Anthony O Gramolini,J Paul Santerre,Vienna Mazzoli

doi:10.1038/s42003-021-02732-8

Jeremy A Antonyshyn, Stefan O P Hofer + Show 5 more

Open Access

https://doi.org/10.1038/s42003-021-02732-8

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Abstract

Endothelial cells are among the fundamental building blocks for vascular tissue engineering. However, a clinically viable source of endothelium has continued to elude the field. Here, we demonstrate the feasibility of sourcing autologous endothelium from human fat – an abundant and uniquely dispensable tissue that can be readily harvested with minimally invasive procedures. We investigate the challenges underlying the overgrowth of human adipose tissue-derived microvascular endothelial cells by stromal cells to facilitate the development of a reliable method for their acquisition. Magnet-assisted cell sorting strategies are established to mitigate the non-specific uptake of immunomagnetic microparticles, enabling the enrichment of endothelial cells to purities that prevent their overgrowth by stromal cells. This work delineates a reliable method for acquiring human adipose tissue-derived microvascular endothelial cells in large quantities with high purities that can be readily applied in future vascular tissue engineering applications.

Highlights

Endothelial cells are among the fundamental building blocks for vascular tissue engineering
Adipose tissue is an attractive source of Endothelial cells (ECs) for vascular tissue engineering because it can be harvested autologously in large quantities with minimally invasive procedures using a cannula coupled to a liposuction pump or even a syringe[10]
Cultures of human adipose tissue-derived microvascular ECs (HAMVECs) were significantly enriched for the CD45–CD31+ immunophenotype when compared with the stromal vascular fraction (98.6 ± 0.9% vs. 0.9 ± 0.6%, respectively; p < 0.0001; Fig. 1b), and they exhibited a characteristic endothelial cobblestone-like morphology (Fig. 1c)

Summary

Introduction

Endothelial cells are among the fundamental building blocks for vascular tissue engineering. While the need for the culture-mediated expansion of human adipose tissue-derived microvascular ECs (HAMVECs) is mitigated by the abundant and uniquely dispensable nature of the tissue, their low prevalence has continued to complicate their acquisition[3,11,12,13,14,15]. Their primary cultures are often overgrown by residual stromal cells from the cell sorting procedure[3,11,12,13,14,15]. There is a clear and unmet need for a readily accessible and non-immunogenic source of endothelium if tissue-engineered constructs are to leave the bench for the bedside

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