Subnormothermic short ‐term cultivation improves the vascularization capacity of adipose tissue‐derived microvascular fragments

In this study we analyzed the effects of normo‐ (37 °C) and subnormothermic (20 °C) short‐term cultivation on their viability and network forming capacity. Ad‐MVF from green fluorescent protein (GFP)+ and GFP‐ C57BL/6 mice were cultivated for 24 h at 37 °C or 20 °C. Freshly isolated, non‐cultivated ad‐MVF served as controls. Number, length, viability, proliferation and angiogenic activity of the ad‐MVF were assessed by microscopic analysis and proteome profiling. GFP+ ad ‐MVF were seeded onto collagen‐glycosaminoglycan matrices, which were implanted into dorsal skinfold chambers of GFP‐ mice to analyze their vascularization by means of intravital fluorescence microscopy, histology and immunohistochemistry. Depending on the temperature short ‐term cultivation of ad‐MVF markedly changed their expression of multiple pro‐ and anti‐angiogenic factors. Moreover, cultivation at 37 °C significantly increased the number of apoptotic cells within ad‐MVF, while 20 °C preserved the viability of ad‐MVF and even promoted the proliferat ion of endothelial and perivascular cells. Accordingly, ad‐MVF cultivated at 20 °C also exhibited an enhanced in vivo vascularization capacity when compared to normothermically cultivated ad‐MVF and non‐cultivated controls. This was indicated by an accelerated network formation, an increased microvascular remodeling and a higher density of GFP+ microvessels within implanted matrices. Thus, if ad ‐MVF require sho...

Source: Journal of Tissue Engineering and Regenerative Medicine - November 23, 2018 Category: Biotechnology Authors: Matthias W. Laschke, Alexander He ß, Claudia Scheuer, Philipp Karschnia, Michael D. Menger Tags: RESEARCH ARTICLE Source Type: research