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Tissue engineering using autologous microcirculatory beds as vascularized bioscaffolds

Edward I. Chang^*,1, Robert G. Bonillas^*,1, Samyra El-ftesi^*, Eric I. Chang^*, Daniel J. Ceradini, Ivan N. Vial^*, Denise A. Chan, Joseph Michaels, V and Geoffrey C. Gurtner^*,2

^* Department of Surgery, Stanford University Medical Center, and

Department of Radiation Biology, Stanford University, Stanford, California, USA; and

Department of Surgery, New York University School of Medicine, New York, New York, USA

²Correspondence: Department of Surgery, Division of Plastic Surgery, 257 Campus Dr., GK-201, Stanford, CA 94305-5148, USA. E-mail: ggurtner{at}stanford.edu

Classic tissue engineering paradigms are limited by the incorporation of a functional vasculature and a reliable means for reimplantation into the host circulation. We have developed a novel approach to overcome these obstacles using autologous explanted microcirculatory beds (EMBs) as bioscaffolds for engineering complex three-dimensional constructs. In this study, EMBs consisting of an afferent artery, capillary beds, efferent vein, and surrounding parenchymal tissue are explanted and maintained for 24 h ex vivo in a bioreactor that preserves EMB viability and function. Given the rapidly advancing field of stem cell biology, EMBs were subsequently seeded with three distinct stem cell populations, multipotent adult progenitor cells (MAPCs), and bone marrow and adipose tissue-derived mesenchymal stem cells (MSCs). We demonstrate MAPCs, as well as MSCs, are able to egress from the microcirculation into the parenchymal space, forming proliferative clusters. Likewise, human adipose tissue-derived MSCs were also found to egress from the vasculature and seed into the EMBs, suggesting feasibility of this technology for clinical applications. We further demonstrate that MSCs can be transfected to express a luciferase protein and continue to remain viable and maintain luciferase expression in vivo. By using the vascular network of EMBs, EMBs can be perfused ex vivo and seeded with stem cells, which can potentially be directed to differentiate into neo-organs or transfected to replace failing organs and deficient proteins.—Chang, E. I., Bonillas, R. G., El-ftesi, S., Chang, E. I., Ceradini, D. J., Vial, I. N., Chan, D. A., Michaels, J., V, Gurtner, G. C. Tissue engineering using autologous microcirculatory beds as vascularized bioscaffolds.

Key Words: stem cells • bioreactor • gene therapy