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Prevascularization of self-organizing engineered heart tissue by human umbilical vein endothelial cells abrogates contractile performance

Cell and Tissue Research volume 350, pages 439–444 (2012)Cite this article

Abstract

Establishing vascularization is a critical obstacle to the generation of engineered heart tissue (EHT) of substantial thickness. Addition of endothelial cells to the formative stages of EHT has been demonstrated to result in prevascularization, or the formation of capillary-like structures. The detailed study of the effects of prevascularization on EHT contractile function is lacking. Here, we evaluated the functional impact of prevascularization by human umbilical vein endothelial cells (HUVECs) in self-organizing EHT. EHT fibers were generated by the self-organization of neonatal rat cardiac cells on a fibrin hydrogel scaffold with or without HUVECs. Contractile function was measured and force–length relationship and rate of force production were assessed. Immunofluorescent studies were used to evaluate arrangement and distribution of HUVECs within the EHT fibers. RT-PCR was used to assess the transcript levels of hypoxia inducible factor-1a (Hif-1α). EHT with HUVECs manifested tubule-like structures at the periphery during fiber formation. After fiber formation, HUVECs were heterogeneously located throughout the EHT fiber and human CD31+ tubule-like structures were identified. The expression level of Hif-1α did not change with the addition of HUVECs. However, maximal force and rate of force generation were not improved in HUVECs containing EHT as compared to control EHT fibers. The addition of HUVECs may result in sparse microvascularization of EHT. However, this perceived benefit is overshadowed by a significant decrease in contractile function and highlights the need for perfused vascularization strategies in order to generate EHT that approaches clinically relevant dimensions.

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References

  • Baar K, Birla R, Boluyt MO, Borschel GH, Arruda EM, Dennis RG (2005) Self-organization of rat cardiac cells into contractile 3-D cardiac tissue. FASEB J 19:275–277

    PubMed  CAS  Google Scholar 

  • Balaban RS (2009) Domestication of the cardiac mitochondrion for energy conversion. J Mol Cell Cardiol 46:832–841

    PubMed  Article  CAS  Google Scholar 

  • Birla RK, Borschel GH, Dennis RG, Brown DL (2005) Myocardial engineering in vivo: formation and characterization of contractile, vascularized three-dimensional cardiac tissue. Tissue Eng 11:803–813

    PubMed  Article  CAS  Google Scholar 

  • Borges J, Mueller MC, Padron NT, Tegtmeier F, Lang EM, Stark GB (2003) Engineered adipose tissue supplied by functional microvessels. Tissue Eng 9:1263–1270

    PubMed  Article  CAS  Google Scholar 

  • Brutsaert DL (2003) Cardiac endothelial-myocardial signaling: its role in cardiac growth, contractile performance, and rhythmicity. Physiol Rev 83:59–115

    PubMed  CAS  Google Scholar 

  • Caspi O, Lesman A, Basevitch Y, Gepstein A, Arbel G, Habib IH, Gepstein L, Levenberg S (2007) Tissue engineering of vascularized cardiac muscle from human embryonic stem cells. Circ Res 100:263–272

    PubMed  Article  CAS  Google Scholar 

  • Chen X, Aledia AS, Popson SA, Him L, Hughes CC, George SC (2010) Rapid anastomosis of endothelial progenitor cell-derived vessels with host vasculature is promoted by a high density of cotransplanted fibroblasts. Tissue Eng A 16:585–594

    Article  CAS  Google Scholar 

  • Huang YC, Khait L, Birla RK (2007) Contractile three-dimensional bioengineered heart muscle for myocardial regeneration. J Biomed Mater Res A 80:719–731

    PubMed  Google Scholar 

  • Ko HC, Milthorpe BK, McFarland CD (2007) Engineering thick tissues—the vascularisation problem. Eur Cells Mater 14:1–18, discussion 18–19

    CAS  Google Scholar 

  • Laschke MW, Mussawy H, Schuler S, Kazakov A, Rucker M, Eglin D, Alini M, Menger MD (2011) Short-term cultivation of in situ prevascularized tissue constructs accelerates inosculation of their preformed microvascular networks after implantation into the host tissue. Tissue Eng A 17:841–853

    Article  CAS  Google Scholar 

  • Lesman A, Habib M, Caspi O, Gepstein A, Arbel G, Levenberg S, Gepstein L (2010) Transplantation of a tissue-engineered human vascularized cardiac muscle. Tissue Eng Part A 16:115–125

    PubMed  Article  CAS  Google Scholar 

  • Leung BM, Sefton MV (2010) A modular approach to cardiac tissue engineering. Tissue Eng Part A 16:3207–3218

    PubMed  Article  CAS  Google Scholar 

  • Li K, Rouleau JL, Andries LJ, Brutsaert DL (1993) Effect of dysfunctional vascular endothelium on myocardial performance in isolated papillary muscles. Circ Res 72:768–777

    PubMed  Article  CAS  Google Scholar 

  • Mathews G, Sondergaard C, Jeffreys A, Childs W, Le BL, Sahota A, Najibi S, Nolta J, Si MS (2012) Computational analysis of contractility in engineered heart tissue. IEEE Trans Biomed Eng 59:1429–1435

    PubMed  Article  Google Scholar 

  • Novosel EC, Kleinhans C, Kluger PJ (2011) Vascularization is the key challenge in tissue engineering. Adv Drug Deliv Rev 63:300–311

    PubMed  Article  CAS  Google Scholar 

  • Olson SD, Kambal A, Pollock K, Mitchell GM, Stewart H, Kalomoiris S, Cary W, Nacey C, Pepper K, Nolta JA (2011) Examination of mesenchymal stem cell-mediated RNAi transfer to Huntington’s disease affected neuronal cells for reduction of huntingtin. Mol Cell Neurosci 49:271–281

    PubMed  Article  Google Scholar 

  • Sekine H, Shimizu T, Hobo K, Sekiya S, Yang J, Yamato M, Kurosawa H, Kobayashi E, Okano T (2008) Endothelial cell coculture within tissue-engineered cardiomyocyte sheets enhances neovascularization and improves cardiac function of ischemic hearts. Circulation 118:S145–S152

    PubMed  Article  CAS  Google Scholar 

  • Sekiya S, Shimizu T, Yamato M, Kikuchi A, Okano T (2006) Bioengineered cardiac cell sheet grafts have intrinsic angiogenic potential. Biochem Biophys Res Commun 341:573–582

    PubMed  Article  CAS  Google Scholar 

  • Sondergaard CS, Hodonsky CJ, Khait L, Shaw J, Sarkar B, Birla R, Bove E, Nolta J, Si MS (2010) Human thymus mesenchymal stromal cells augment force production in self-organized cardiac tissue. Ann Thorac Surg 90:796–803, discussion 803–794

    PubMed  Article  Google Scholar 

  • Stevens KR, Kreutziger KL, Dupras SK, Korte FS, Regnier M, Muskheli V, Nourse MB, Bendixen K, Reinecke H, Murry CE (2009) Physiological function and transplantation of scaffold-free and vascularized human cardiac muscle tissue. Proc Natl Acad Sci USA 106:16568–16573

    PubMed  Article  CAS  Google Scholar 

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Acknowledgments

This work was supported in part by the Children Miracle Network Young Investigator Award (MSI) and the UC Davis School of Medicine. C.S.S. was in part supported by a training grant from the California Institute for Regenerative Cures. We acknowledge S. Taylor for providing statistics consultation.

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Affiliations

  1. Department of Cardiothoracic Surgery, University of California at Davis Medical Center, Sacramento, CA, 95817, USA

    Claus Svane Sondergaard, Russell Witt, Grant Mathews, Skender Najibi, Lisa Le & Tracy Clift

  2. Institute for Regenerative Cures, University of California at Davis Medical Center, Sacramento, CA, USA

    Claus Svane Sondergaard & Ming-Sing Si

  3. Department of Cardiac Surgery, University of Michigan, 11-735 C.S. Mott Children’s Hospital 1540 E. Hospital Drive, SPC 4204, Ann Arbor, MI, 48109, USA

    Ming-Sing Si

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  1. Claus Svane Sondergaard
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  2. Russell Witt
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  3. Grant Mathews
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  4. Skender Najibi
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  5. Lisa Le
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  6. Tracy Clift
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  7. Ming-Sing Si
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Correspondence to Ming-Sing Si.

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Sondergaard, C.S., Witt, R., Mathews, G. et al. Prevascularization of self-organizing engineered heart tissue by human umbilical vein endothelial cells abrogates contractile performance. Cell Tissue Res 350, 439–444 (2012). https://doi.org/10.1007/s00441-012-1492-7

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  • DOI: https://doi.org/10.1007/s00441-012-1492-7

Keywords

  • Engineered heart tissue
  • Prevascularization
  • Tissue engineering
  • HUVECs