In vitro fabrication of autologous living tissue-engineered vascular grafts based on prenatally harvested ovine amniotic fluid-derived stem cells

• Published in: Journal of tissue engineering and regenerative medicine Vol. 10; no. 1; pp. 52 - 70
• Main Authors: Weber, Benedikt, Kehl, Debora, Bleul, Ulrich, Behr, Luc, Sammut, Sébastien, Frese, Laura, Ksiazek, Agnieszka, Achermann, Josef, Stranzinger, Gerald, Robert, Jérôme, Sanders, Bart, Sidler, Michele, Brokopp, Chad E., Proulx, Steven T., Frauenfelder, Thomas, Schoenauer, Roman, Emmert, Maximilian Y., Falk, Volkmar, Hoerstrup, Simon P.
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01-01-2016; Hindawi Limited
• Subjects: AC133 Antigen; Amnion - cytology; amniotic fluid; Amniotic Fluid - cytology; Animals; Antigens, CD - metabolism; Blood Vessel Prosthesis; cardiovascular tissue engineering; Cell Differentiation; Cell Proliferation; Cell Separation; Cell Shape; Cell Survival; Chromosome Aberrations; Endothelial Cells - cytology; Female; fetal model; Genotype; Glycoproteins - metabolism; Karyotyping; mesenchymal stem cells; Mesenchymal Stromal Cells; Myofibroblasts - cytology; ovine in-vivo model; Peptides - metabolism; Phenotype; Pregnancy; Regenerative medicine; Sheep; Tissue engineering; Tissue Engineering - methods; Tissue Scaffolds - chemistry; Transplantation, Autologous; vascular graft; cardiovascular tissue engineering; fetal model; vascular graft; mesenchymal stem cells; ovine in-vivo model; amniotic fluid
• ISSN: 1932-6254; 1932-7005
• DOI: 10.1002/term.1781

Amniotic fluid cells (AFCs) have been proposed as a valuable source for tissue engineering and regenerative medicine. However, before clinical implementation, rigorous evaluation of this cell source in clinically relevant animal models accepted by regulatory authorities is indispensable. Today, the ovine model represents one of the most accepted preclinical animal models, in particular for cardiovascular applications. Here, we investigate the isolation and use of autologous ovine AFCs as cell source for cardiovascular tissue engineering applications. Fetal fluids were aspirated in vivo from pregnant ewes (n = 9) and from explanted uteri post mortem at different gestational ages (n = 91). Amniotic non‐allantoic fluid nature was evaluated biochemically and in vivo samples were compared with post mortem reference samples. Isolated cells revealed an immunohistochemical phenotype similar to ovine bone marrow‐derived mesenchymal stem cells (MSCs) and showed expression of stem cell factors described for embryonic stem cells, such as NANOG and STAT‐3. Isolated ovine amniotic fluid‐derived MSCs were screened for numeric chromosomal aberrations and successfully differentiated into several mesodermal phenotypes. Myofibroblastic ovine AFC lineages were then successfully used for the in vitro fabrication of small‐ and large‐diameter tissue‐engineered vascular grafts (n = 10) and cardiovascular patches (n = 34), laying the foundation for the use of this relevant pre‐clinical in vivo assessment model for future amniotic fluid cell‐based therapeutic applications. Copyright © 2013 John Wiley & Sons, Ltd.