title: Adipose-derived perivascular mesenchymal stromal/stem cells promote functional vascular tissue engineering for cardiac regenerative purposes.
abstract: Cardiac tissue engineering approaches have the potential to regenerate functional myocardium with intrinsic vascular networks. This study compared the relative effects of human adipose-derived stem/stromal cells (hASCs) and human dermal fibroblasts (hDFs) in cocultures with neonatal rat ventricular cardiomyocytes (NRVCMs) and human umbilical vein endothelial cells (HUVECs). At the same ratios of NRVCM:hASC and NRVCM:hDF, the hASC cocultures displayed shorter action potentials and maintained capture at faster pacing rates. Similarly, in coculture with HUVECs, hASC:HUVEC exhibited superior ability to support vascular capillary network formation relative to hDF:HUVEC. Based on these studies, a range of suitable cell ratios were determined to develop a triculture system. Six seeding ratios of NRVCM:hASC:HUVEC were tested and it was found that a ratio of 500:50:25 cells (i.e. 250,000:25,000:12,500 cells/cm2) resulted in the formation of robust vascular networks while retaining action potential durations and propagation similar to pure NRVCM cultures. Tricultures in this ratio exhibited an average conduction velocity of 20 ± 2 cm/s, action potential durations at 80% repolarization (APD80) and APD30of 122 ± 5 ms and 59 ± 4 ms, respectively, and maximum capture rate of 7.4 ± 0.6 Hz. The NRVCM control groups had APD80and APD30of 120 ± 9 ms and 51 ± 5 ms, with a maximum capture rate of 7.3 ± 0.2 Hz. In summary, the combination of hASCs in the appropriate ratios with NRVCMs and HUVECs can facilitate the formation of densely vascularized cardiac tissues that appear not to impact the electrophysiological function of cardiomyocytes negatively. Copyright © 2017 John Wiley & Sons, Ltd.
authors.affiliation: Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA.|Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Morrissette-McAlmon
authors.affiliation: Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Adriana
authors.initials: A
authors.lastname: Blazeski
authors.affiliation: Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA.|Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Somers
authors.affiliation: Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Geran
authors.initials: G
authors.lastname: Kostecki
authors.affiliation: Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Leslie
authors.initials: L
authors.lastname: Tung
authors.affiliation: Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA.|Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.|Department of Material Sciences & Engineering, Johns Hopkins University, School of Engineering, Baltimore, MD, USA.|Institute for Nanobiotechnology (INBT), Johns Hopkins University School of Engineering, Baltimore, MD, USA.
authors.forename: Warren L
authors.identifier: 0000-0001-6099-6469
authors.initials: WL
authors.lastname: Grayson
mesh_terms: D000818:Animals; D001807:Blood Vessel Prosthesis; D055724:Electrophysiological Phenomena; D005260:Female; D006321:Heart / Q000502:physiology*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008875:Middle Aged; D032383:Myocytes, Cardiac / Q000166:cytology; D018919:Neovascularization, Physiologic; D017207:Rats, Sprague-Dawley; D012038:Regeneration / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*
keywords: Adipose-derived stem cells; Cardiac tissue engineering; Neonatal rat ventricular cardiomyocytes; dermal fibroblasts; electrophysiology; vascularized grafts