## 5 Conclusions and Further Perspectives  
In this study, we have prepared decellularized pericardium repopulated with adipose tissue-derived stromal cells for potential use as implantable cardiovascular patches. To achieve this aim, novel optimized dynamic decellularization and recellularization systems, fabricated in our laboratory, have been used and demonstrated in this study. In the decellularization system, the planar tissue is exposed on both sides to a continuous flow of the decellularization agents (SDS and DNAse) and the washing medium (dH\({}_{2}\)O), which are automatically changed. This process, taking approximately 1 day, provided a completely decellularized non-cytotoxic matrix suitable for further recellularization. The recellularization was performed using porcine adipose tissue-derived stromal cells and a unique custom-made dynamic bioreactor generating pulsatile pressure stress. This mechanical stimulation markedly accelerated the penetration of cells inside the decellularized tissue, their proliferation, and particularly their differentiation towards smooth muscle cells, i.e., an important cell type and the most numerous cell type in the vascular wall. While the decellularization and recellularization procedures in conventional static systems usually take several weeks, our newly developed systems need only about two weeks, including decellularization (approx. 1 day), isolation, and expansion of ASCs (8-10 days) and recellularization (5 days). The newly prepared pericardium patches recellularized with pre-differentiated stromal cells are intended for treating an experimentally-induced carotid artery defect in a porcine model in vivo in order to develop novel cardiovascular patches that can be applied in human medicine.  
Conceptualization R.M., M.K., J.C., S.F.; Data curation [PERSON]; Formal analysis [PERSON], [PERSON]; Funding acquisition [PERSON], [PERSON], S.F., J.P.; Investigation [PERSON], M.K., J.C., S.P., A.B.; Methodology [PERSON], J.C., R.M., S.F.; Project administration [PERSON], P.K.; Resources J.L., Z.S., I.M., D.T.,; Software [PERSON], J.S.; Supervision [PERSON], J.R.; Validation [PERSON], S.P.; Visualization [PERSON], [PERSON]; Writing--original draft [PERSON], M.K., [PERSON]; Writing--review & editing [PERSON], J.R., L.B. All authors have read and agreed to the published version of the manuscript.  
The research was supported by the Ministry of Health of the Czech Republic grants No. NV18-02-00422 and 15-29153A; by the Ministry of Education, Youth and Sports of CR by the project \"BIOCEV\" reg. no. CZ.105/1.100/02.0109. All rights reserved.  
The authors thank [PERSON] (Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic) for her assistance with the preparation of thin cryosections of samples; [PERSON] (Institute of Clinical and Experimental Medicine, Prague, Czech Republic) caring for experimental animals and staff of Steinhauser s.r.o. for allowing the porcine pericardium to be extracted. [PERSON] (Czech Technical University, Prague) is gratefully acknowledged for his language revision of the manuscript.  
The authors declare no conflict of interest.  
The animal experiments in this study were approved by the Ministry of Health of the Czech Republic reference no MZDZ 23132/2018-4/OVZ approval no 37/2018 and reference no MZDZ 48434/2015-OVZ-30.0-21.8.15 approval no 58/2015. A minimal number of animals was used. All procedures described were done under general anaesthesia and according to ethical guidelines to minimize the pain and discomfort of animals. The Institute of Clinical and Experimental has authorized facilities and fully equipped operating theatres for performing these animal experiments.