This protocol describes detailed practical procedures for generating 3D intact and perfusable microvascular network that connects to microfluidic channels without appreciable leakage. This advanced 3D microvascular network model incorporates different stages of vascular development including vasculogenesis, endothelial cell (EC) lining, sprouting angiogenesis, and anastomosis in sequential order. The capillary network is first induced via vasculogenesis in a middle tissue chamber and then EC linings along the microfluidic channel on either side serve as artery and vein. The anastomosis is then induced by sprouting angiogenesis to facilitate tight interconnection between the artery/vein and the capillary network. This versatile device design and its robust construction methodology establish a physiological microcirculation transport model of interconnected perfused vessels from artery to vascularized tissue to vein.
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
This work was supported by grants from the National Institutes of Health: UH3 TR00048 and PQD5 CA180122. C.C.W.H. receives support from the Chao Family Comprehensive Cancer Center (CFCCC) through an NCI Center Grant award P30A062203. X.W. receives support from National Natural Science Foundation of China (No. 31600781). We would also like to thank the permission of The Royal Society of Chemistry (RSC) for reproduction of materials from Lab on a Chip journal.
Lee H, Chung M, Jeon NL (2014) Microvasculature: an essential component for organ-on-chip systems. MRS Bull 39(1):51–59CrossRefGoogle Scholar
Schimek K, Busek M, Brincker S et al (2013) Integrating biological vasculature into a multi-organ-chip microsystem. Lab Chip 13(18):3588–3598CrossRefPubMedGoogle Scholar
Esch MB, Post DJ, Shuler ML et al (2011) Characterization of in vitro endothelial linings grown within microfluidic channels. Tissue Eng A 17(23–24):2965–2971CrossRefGoogle Scholar
Bischel LL, Young EWK, Mader BR et al (2013) Tubeless microfluidic angiogenesis assay with three-dimensional endothelial-lined microvessels. Biomaterials 34(5):1471–1477CrossRefPubMedGoogle Scholar
Booth R, Noh S, Kim H (2014) A multiple-channel, multiple-assay platform for characterization of full-range shear stress effects on vascular endothelial cells. Lab Chip 14(11):1880–1890CrossRefPubMedGoogle Scholar
Lee H, Kim S, Chung M et al (2014) A bioengineered array of 3D microvessels for vascular permeability assay. Microvasc Res 91:90–98CrossRefPubMedGoogle Scholar
Kim S, Lee H, Chung M et al (2013) Engineering of functional, perfusable 3D microvascular networks on a chip. Lab Chip 13(8):1489–1500CrossRefPubMedGoogle Scholar
Yeon JH, Ryu HR, Chung M et al (2012) In vitro formation and characterization of a perfusable three-dimensional tubular capillary network in microfluidic devices. Lab Chip 12(16):2815–2822CrossRefPubMedGoogle Scholar
Vickerman V, Blundo J, Chung S et al (2008) Design, fabrication and implementation of a novel multi-parameter control microfluidic platform for three-dimensional cell culture and real-time imaging. Lab Chip 8(9):1468–1477CrossRefPubMedPubMedCentralGoogle Scholar
Young EWK (2013) Advances in microfluidic cell culture systems for studying angiogenesis. J Lab Autom 18(6):427–436CrossRefPubMedGoogle Scholar
Chiu LL, Montgomery M, Liang Y et al (2012) Perfusable branching microvessel bed for vascularization of engineered tissues. Proc Nat Acad Sci U S A 109(50):E3414–E3423CrossRefGoogle Scholar
Whisler JA, Chen MB, Kamm RD (2014) Control of perfusable microvascular network morphology using a multiculture microfluidic system. Tissue Eng 20(7):543–552CrossRefGoogle Scholar
Diaz-Santana A, Shan M, Stroock AD (2015) Endothelial cell dynamics during anastomosis in vitro. Integr Biol 7(4):454–466CrossRefGoogle Scholar
Huh D, Torisawa YS, Hamilton GA et al (2012) Microengineered physiological biomimicry: organs-on-chips. Lab Chip 12(12):2156–2164CrossRefPubMedGoogle Scholar
Hsu YH, Moya ML, Hughes CCW et al (2013) Full range physiological mass transport control in 3D tissue cultures. Lab Chip 13(1):81–89CrossRefPubMedGoogle Scholar
Hsu YH, Moya ML, Hughes CCW et al (2013) A microfluidic platform for generating large-scale nearly identical human microphysiological system arrays. Lab Chip 13(15):2990–2998CrossRefPubMedPubMedCentralGoogle Scholar
Wang X, Phan DTT, Sobrino A et al (2016) Engineering anastomosis between living capillary networks and endothelial cell-lined microfluidic channels. Lab Chip 16(2):282–290CrossRefPubMedPubMedCentralGoogle Scholar
Wang X, Phan DTT, Zhao D et al (2016) An on-chip microfluidic pressure regulator that facilitates reproducible loading of cells and hydrogels into microphysiological system platforms. Lab Chip 16(5):868–876CrossRefPubMedPubMedCentralGoogle Scholar