Thursday September 15
| Short communications
Live human arterial tissue slices for bench top research on pathophysiology of aortic aneurysms; up to 90 days ex vivo preservation Jorn P. Meekel, René J.P. Musters, Behrouz Z. Doulabi, Dimitra Micha, Gerard Pals, Willem Wisselink, Jan D. Blankensteijn, Kak K. Yeung VU Medical Center, Amsterdam, The Netherlands
Currently, research on abdominal aortic aneurysm (AAA) pathophysiology is limited to fixated cells or isolated cell cultures1,2,3. However, due to these techniques, cell-cell interactions and communication with extracellular matrix are lost. These interactions are likely to play a role in the AAA pathogenesis4,5,6,7. We hereby present an innovative method to cut live vascular tissue slices and preserve cell viability and cellular organization for research into AAA in which anatomical, functional and microstructural characteristics are of interest. Peroperatively harvested tissue was transported in ice-cold Custodiol and cut into cubes. Subsequently, cubes were submerged in agarose, glued on an anvil, and cut into slices of 150 μm. Slices were cultured at 37°Celsius in media supplemented with antibiotics. Viability analysis was performed up to 92 days after harvesting using LIVE/DEAD® Viability/ Cytotoxicity Kit. Cell type characterization was achieved by staining for CD45, CD68, α-SMA/ smoothelin to identify leukocytes, macrophages and fibroblasts/smooth muscle cells (SMC) respectively. Additionally, tissues were digested using collagenase to study individual cells and analyze cellular populations in live tissues. Tissue slice analysis showed a stable viability of 40% until 7 days (graph. 1) and after improvement of the protocol up to 92 days after harvesting (with outgrowth of new cells). Live cells were mainly seen centrally in the tissue, while dead cells were observed at cutting edges. Live cells were differentiated by qualitative analysis based on cell morphology and specific marker expression. The majority of studied live cells were fibroblasts/SMC. Furthermore, leukocytes and macrophages were observed. These findings were in accordance with the findings in cells of digested tissues. Vitality and organization of tissue sections of aneurysmal and non-aneurysmal vascular tissue can be preserved until 92 days after harvesting. This study provides a solid base for further experimental research on pathophysiological mechanisms underlying aneurysms and possibly other vascular diseases.
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IMAD 2016, September 15-17
IMAD