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Ex Vivo Human Aortic Punch Tissue: A New Approach for Investigating Angiogenesis in Cardiovascular Diseases
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DOI：10.17576/jsm-2026-5504-08.png)
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Abstract
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Cardiovascular diseases (CVDs) remain the leading cause of mortality worldwide, with pathological angiogenesis contributing to disease progression. Conventional ex vivo angiogenesis assays often employ animal tissues, but physiological differences limit translational relevance. This study establishes a human ex vivo angiogenesis model using discarded aortic punch tissues from coronary artery bypass graft (CABG) surgeries, providing a clinically relevant platform for investigating angiogenic mechanisms. Two culture approaches of embedded and sandwich techniques were optimized using aortic punch tissues maintained in Matrigel for up to 27 days. Angiogenic sprouting was assessed microscopically, quantified with ImageJ, and analysed using repeated measure ANOVA with Bonferroni post hoc test. No sprouting was observed with the embedded method. In contrast, the sandwich method induced vessel sprouting, initiating at Day 7 with progressive increases in sprouting area and branching length through Day 27 (p < 0.01). Under hypoxic conditions (1% O-2), sprouting was significantly enhanced compared to normoxia (20% O-2), with greater sprouting area, branch number, and branching length. Hypoxia also elevated HIF-1 alpha expression, particularly at Day 7 and peaked at Day 14 (p < 0.05) compare to normoxia, confirming hypoxia-driven angiogenic pathways. The sandwich culture method effectively supports angiogenesis in human aortic punch tissues, unlike the embedded approach. Hypoxia further amplifies angiogenic responses via HIF-1 alpha signalling. This optimized human ex vivo assay offers a clinically relevant model bridging in vitro and animalbased systems, serving as a valuable tool for mechanistic studies and therapeutic exploration in cardiovascular disease.
Keywords:
Angiogenesis
aortic ring assay
ex vivo
human aortic tissue
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