In vitro pre-vascularization strategies for tissue engineered constructs – Bioprinting and others

VIEWS - 314 (Abstract) 286 (PDF)
Andy Wen Loong Liew, Yilei Zhang


Tissue-engineered products commercially available today have been limited to thin avascular tissue such as skin and cartilage. The fabrication of thicker, more complex tissue still eludes scientists today. One reason for this is the lack of effective techniques to incorporate functional vascular networks within thick tissue constructs. Vascular networks provide cells throughout the tissue with adequate oxygen and nutrients; cells located within thick un-vascularized tissue implants eventually die due to oxygen and nutrient deficiency. Vascularization has been identified as one of the key components in the field of tissue engineering. In order to fabricate biomimetic tissue which accurately recapitulates our native tissue environment, in vitro pre-vascularization strategies need to be developed. In this review, we describe various in vitro vascularization techniques developed recently which employ different technologies such as bioprinting, microfluidics, micropatterning, wire molding, and cell sheet engineering. We describe the fabrication process and unique characteristics of each technique, as well as provide our perspective on the future of the field.


Pre-vascularization; bio-printing; endothelial; tissue engineering

Full Text:



U.S. Department of Health & Human Services, 2015, Organ donation statistics, viewed on October 2, 2016,

Langer R and Vacanti J P, 1993, Tissue engineering. Science, vol.260(5110): 920–926.

Hüsing B, Bührlen B, Gaisser S, et al. 2003, Human tissue-engineered products: today’s markets and future prospects. Science And Technology: 1–58.

Sakaguchi K, Shimizu T and Okano T, 2015, Construction of three-dimensional vascularized cardiac tissue with cell sheet engineering. Journal of Controlled Release, vol.205: 83–88.

Yeong W Y, Sudarmadji N, Yu H Y, et al. 2010, Porous polycaprolactone scaffold for cardiac tissue engineering fabricated by selective laser sintering. Acta Biomaterialia, vol.6(6): 2028–2034.

Lee P J, Hung P J and Lee L P, 2007, An artificial liver sinusoid with a microfluidic endothelial-like barrier for primary hepatocyte culture. Biotechnology and Bioengineering, vol.97(5): 1340–1346.

Tonsomboon K and Oyen M L, 2013, Composite electrospun gelatin fiber-alginate gel scaffolds for mechanically robust tissue engineered cornea. Journal of the Mechanical Behavior of Biomedical Materials, vol.21: 185–194.

Jungebluth P, Alici E, Baiguera S, et al. 2011, Tracheobronchial transplantation with a stem-cell-seeded bioartificial nanocomposite: a proof-of-concept study. The Lancet, vol.378(9808): 1997–2004.

Hasan A, Memic A, Annabi N, et al. 2014, Electrospun scaffolds for tissue engineering of vascular grafts. Acta Biomaterialia, vol.10(1): 11–25.

Atala A, Bauer S B, Soker S, et al. 2006, Tissue-engineered autologous bladders for patients needing cystoplasty. Lancet, vol.367(9518): 1241–1246.

Folkman J and Hochberg M, 1973, Self-regulation of growth in three dimensions. The Journal of Experimental Medicine, vol.138(4): 745–753.

Rouwkema J, Rivron N C and van Blitterswijk C A, 2008, Vascularization in tissue engineering. Trends in Biotechnology, vol.26(8): 434–441.

Schmeichel K L and Bissell M J, 2010, Modeling tissue-specific signaling and organ function in three dimensions. Journal of Cell Science, vol.116(Pt 12): 2377–2388.

Griffith L G and Swartz M A, 2006, Capturing complex 3D tissue physiology in vitro. Nature Reviews: Molecular Cell Biology, vol.7(3): 211–224.

Auger F A, Gibot L and Lacroix D, 2013, The pivotal role of vascularization in tissue engineering. Annual Review of Biomedical Engineering, vol.15: 177–200.

Baranski J D, Chaturvedi R R, Stevens K R, et al. 2013, Geometric control of vascular networks to enhance engineered tissue integration and function. Proceedings of the National Academy of Sciences of the United States of America, vol.110(19): 7586–7591.

Krewski D, Acosta Jr D, Andersen M, et al. 2010, Toxicity Testing in the 21st Century: a vision and a strategy. Journal of Toxicology and Environmental Health. Part B: Critical Reviews, vol.13(2–4), 51–138.

Elliott N T and Yuan F, 2011, A review of three-dimensional in vitro tissue models for drug discovery and transport studies. Journal of Pharmaceutical Sciences, vol.100: 59–74.

Naito H, Yoshimura M, Mizuno T, et al. 2013, The advantages of three-dimensional culture in a collagen hydrogel for stem cell differentiation. Journal of Biomedical Materials Research. Part A, vol.101(10): 2838–2845.

Baker B M and Chen C S., 2012, Deconstructing the third dimension — how 3D culture microenvironments alter cellular cues. Journal of Cell Science, vol.125(13): 3015–3024.

Soucy P A and Romer L H, 2009, Endothelial cell adhesion, signaling, and morphogenesis in fibroblast-derived matrix. Matrix Biology, vol.28(5): 273–283.

Wang R, Xu J, Juliette L, et al. 2005, Three-dimensional co-culture models to study prostate cancer growth, progression, and metastasis to bone. Seminars in Cancer Biology, vol.15(5): 353–364.

Burke A S, MacMillan-Crow L A and Hinson J A, 2010, The hepatocyte suspension assay is superior to the cultured hepatocyte assay for determining mechanisms of acetaminophen hepatotoxicity relevant to in vivo toxicity. Chemical Research in Toxicology, vol.23(12): 1855–1858.

Mingoia R T, Nabb D L, Yang C-H, et al. 2007, Primary culture of rat hepatocytes in 96-well plates: effects of extracellular matrix configuration on cytochrome P450 enzyme activity and inducibility, and its application in in vitro cytotoxicity screening. Toxicology In Vitro, vol.21(1): 165–173.

Zhang W, Wray L S, Rnjak-Kovacina J, et al. 2015, Vascularization of hollow channel-modified porous silk scaffolds with endothelial cells for tissue regeneration. Biomaterials, vol.56: 68–77.

Mitchell G M and Morrison W A, 2014, In vivo vascularization for large-volume soft tissue-engineering. Vascularization — Regenerative Medicine and Tissue Engineering: 343–362.

Dababneh A B and Ozbolat I T, 2014, Bioprinting technology: a current state-of-the-art review. Journal of Manufacturing Science and Engineering, vol.136(6): 61016.

Hinton T J, Jallerat Q, Palchesko R N, et al. 2015, Three-dimensional printing of complex biological structures by freeform reversible embedding of suspended hydrogels. Science Advances, vol.1(9): e1500758.

Schuurman W, Khristov V, Pot M W, et al. 2011, Bioprinting of hybrid tissue constructs with tailorable mechanical properties. Biofabrication, vol.3(2): 21001.

Norotte C, Marga F S, Niklason L E, et al. 2009, Scaffold-free vascular tissue engineering using bioprinting. Biomaterials, vol.30(30): 5910–5917.

Tan Y, Richards D J, Trusk T C, et al. 2014, 3D printing facilitated scaffold-free tissue unit fabrication. Biofabrication, vol.6(2): 24111.

Miller J S, Stevens K R, Yang M T, et al. 2012, Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues. Nature Materials, vol.11(9): 768–774.

Ovsianikov A, Gruene M, Pflaum M, et al. 2010, Laser printing of cells into 3D scaffolds. Biofabrication, vol.2(1): 14104.

Ringeisen B R, Othon C M, Barron J A, et al. 2006, Jet-based methods to print living cells. Biotechnology Journal, vol.1(9): 930–948.

Lin H, Zhang D, Alexander P G, et al. 2013, Application of visible light-based projection stereolithography for live cell-scaffold fabrication with designed architecture. Biomaterials, vol.34(2): 331–339.

Chan V, Zorlutuna P, Jeong J H, et al. 2010, Three-dimensional photopatterning of hydrogels using stereolithography for long-term cell encapsulation. Lab on a Chip, vol.10(16): 2062–2070.

Kolesky D B, Truby R L, Gladman A S, et al. 2014, 3D bioprinting of vascularized, heterogeneous cell-laden tissue constructs. Advanced Materials, vol.26(19): 3124–3130.

Jia W, Gungor-Ozkerim P S, Zhang Y S, et al. 2016, Direct 3D bioprinting of perfusable vascular constructs using a blend bioink. Biomaterials, vol.106: 58–68.

Bertassoni L E, Cecconi M, Manoharan V, et al. 2014, Hydrogel bioprinted microchannel networks for vascularization of tissue engineering constructs. Lab on a Chip, vol.14(13): 2202–2211.

Van Der Meer A D, Poot A A, Duits M H G, et al. 2009, Microfluidic technology in vascular research. Journal of Biomedicine and Biotechnology, vol.2009: 823148.

Zheng Y, Chen J, Craven M, et al. 2012, In vitro microvessels for the study of angiogenesis and thrombosis. Proceedings of the National Academy of Sciences of the United States of America, vol.109(24): 9342–9347.

Tocchio A, Tamplenizza M, Martello F, et al. 2015, Versatile fabrication of vascularizable scaffolds for large tissue engineering in bioreactor. Biomaterials, vol.45: 124–131.

Song J W, Bazou D and Munn L L, 2012, Anastomosis of endothelial sprouts forms new vessels in a tissue analogue of angiogenesis. Integrative Biology, vol.4(8): 857–862.

Kim S, Lee H, Chung M, et al. 2013, Engineering of functional, perfusable 3D microvascular networks on a chip. Lab on a Chip, vol.13: 1489–1500. http://dx.doi/org/10.1039/c3lc41320a

Moya M L, Hsu Y-H, Lee A P, et al. 2013, In vitro perfused human capillary networks. Tissue Engineering Part C: Methods, vol.19(9): 730–737.

Chiu L L Y, Montgomery M, Liang Y, et al. 2012, Perfusable branching microvessel bed for vascularization of engineered tissues. Proceedings of the National Academy of Sciences of the United States of America, vol.109(50): E3414–23.

Yeon J H, Ryu H R, Chung M, et al. 2012, In vitro formation and characterization of a perfusable three-dimensional tubular capillary network in microfluidic devices. Lab on a Chip, vol.12(16): 2815.

Jakab K, Norotte C, Marga F, et al. 2010, Tissue engineering by self-assembly and bio-printing of living cells. Biofabrication, vol.2(2): 22001.

Dickinson L E, Moura M E and Gerecht S, 2010, Guiding endothelial progenitor cell tube formation using patterned fibronectin surfaces. Soft Matter, vol.6(20): 5109.

Raghavan S, Nelson C M, Baranski J D, et al. 2010, Geometrically controlled endothelial tubulogenesis in micropatterned gels. Tissue engineering. Part A, vol.16(7): 2255–2263.

Chaturvedi R R, Stevens K R, Solorzano R D, et al. 2015, Patterning vascular networks in vivo for tissue engineering applications. Tissue Engineering Part C: Methods, vol.21(5): 509–517.

Aubin H, Nichol J W, Hutson C B, et al. 2010, Directed 3D cell alignment and elongation in microengineered hydrogels. Biomaterials, vol.31(27): 6941–6951.

Nikkhah M, Eshak N, Zorlutuna P, et al. 2012, Directed endothelial cell morphogenesis in micropatterned gelatin methacrylate hydrogels. Biomaterials, vol.33(35): 9009–9018.

van der Meer A D, Orlova V V, ten Dijke P, et al. 2013, Three-dimensional co-cultures of human endothelial cells and embryonic stem cell-derived pericytes inside a microfluidic device. Lab on a Chip, vol.13(18): 3562–3568.

Leslie-Barbick J E, Shen C, Chen C, et al. 2011, Micron-scale spatially patterned, covalently immobilized vascular endothelial growth factor on hydrogels accelerates endothelial tubulogenesis and increases cellular angiogenic responses. Tissue engineering. Part A, vol.17(1–2): 221–229.

Nichol J W, Koshy S T, Bae H, et al. 2010, Cell-laden microengineered gelatin methacrylate hydrogels. Biomaterials, vol.31(21): 5536–5544.

Park J H, Chung B G, Lee W G, et al. 2010, Microporous cell-laden hydrogels for engineered tissue constructs. Biotechnology and Bioengineering, vol.106(1): 138–148.

Yao L, de Ruiter G C W, Wang H, et al. 2010, Controlling dispersion of axonal regeneration using a multichannel collagen nerve conduit. Biomaterials, vol.31(22): 5789–5797.

Chrobak K M, Potter D R and Tien J, 2006, Formation of perfused, functional microvascular tubes in vitro. Microvascular Research, vol.71(3): 185–196.

Sadr N, Zhu M, Osaki T, et al. 2011, SAM-based cell transfer to photopatterned hydrogels for microengineering vascular-like structures. Biomaterials, vol.32(30): 7479–7490.

Yoshida H, Matsusaki M and Akashi M, 2013, Multilayered blood capillary analogs in biodegradable hydrogels for in vitro drug permeability assays. Advanced Functional Materials, vol.23(14): 1736–1742.

Price G M, Wong K H K, Truslow J G, et al. 2010, Effect of mechanical factors on the function of engineered human blood microvessels in microfluidic collagen gels. Biomaterials, vol.31(24): 6182–6189.

Nishida K, Yamato M, Hayashida Y, et al. 2004, Functional bioengineered corneal epithelial sheet grafts from corneal stem cells expanded ex vivo on a temperature-responsive cell culture surface. Transplantation, vol.77(3): 379–385.

Yamato M, Utsumi M, Kushida A, et al. 2001, Thermo-responsive culture dishes allow the intact harvest of multilayered keratinocyte sheets without dispase by reducing temperature. Tissue Engineering, vol.7(4): 473–480.

Shimizu T, Yamato M, Kikuchi A, et al. 2003, Cell sheet engineering for myocardial tissue reconstruction. Biomaterials, vol.24(13): 2309–2316.

Asakawa N, Shimizu T, Tsuda Y, et al. 2010, Pre-vascularization of in vitro three-dimensional tissues created by cell sheet engineering. Biomaterials, vol.31(14): 3903–3909.

Muraoka M, Shimizu T, Itoga K, et al. 2013, Control of the formation of vascular networks in 3D tissue engineered constructs. Biomaterials, vol.34(3): 696–703.

Shimizu T, Sekine H, Yang J, et al. 2006, Polysurgery of cell sheet grafts overcomes diffusion limits to produce thick, vascularized myocardial tissues. The FASEB Journal, vol.20(6): 1–20.

Sakaguchi K, Shimizu T, Horaguchi S, et al. 2013, In vitro engineering of vascularized tissue surrogates. Scientific Reports, vol.3: 1316.

Sekine H, Shimizu T, Hobo K, et al. 2008, Endothelial cell coculture within tissue-engineered cardiomyocyte sheets enhances neovascularization and improves cardiac function of ischemic hearts. Circulation, vol.118(14 Suppl): 145–153.

Wong H K, Ivan Lam C R, Wen F, et al. 2016, Novel method to improve vascularization of tissue engineered constructs with biodegradable fibers. Biofabrication, vol.8(1): 15004.

Nishiguchi A, Yoshida H, Matsusaki M, et al. 2011, Rapid construction of three-dimensional multilayered tissues with endothelial tube networks by the cell-accumulation technique. Advanced Materials, vol.23(31): 3506–3510.



  • There are currently no refbacks.

Copyright (c) 2017 Andy Wen Loong Liew, Yilei Zhang

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Recent Articles | About Journal | For Author | Fees | About Whioce

Copyright © Whioce Publishing Pte Ltd. All Rights Reserved.