Advertisement

3D Anastomosed Microvascular Network Model with Living Capillary Networks and Endothelial Cell-Lined Microfluidic Channels

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1612)

Abstract

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.

Key words

3D microvascular network Microfluidic chip Vasculogenesis EC lining Sprouting angiogenesis Anastomosis Non-physiological leakage Organ-on-a-chip 

Notes

Acknowledgments

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.

References

  1. 1.
    Lee H, Chung M, Jeon NL (2014) Microvasculature: an essential component for organ-on-chip systems. MRS Bull 39(1):51–59CrossRefGoogle Scholar
  2. 2.
    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
  3. 3.
    Hasan A, Paul A, Vrana NE et al (2014) Microfluidic techniques for development of 3D vascularized tissue. Biomaterials 35(26):7308–7325CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    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
  5. 5.
    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
  6. 6.
    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
  7. 7.
    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
  8. 8.
    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
  9. 9.
    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
  10. 10.
    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
  11. 11.
    Young EWK (2013) Advances in microfluidic cell culture systems for studying angiogenesis. J Lab Autom 18(6):427–436CrossRefPubMedGoogle Scholar
  12. 12.
    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
  13. 13.
    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
  14. 14.
    Diaz-Santana A, Shan M, Stroock AD (2015) Endothelial cell dynamics during anastomosis in vitro. Integr Biol 7(4):454–466CrossRefGoogle Scholar
  15. 15.
    Chan CY, Huang PH, Guo F et al (2013) Accelerating drug discovery via organs-on-chips. Lab Chip 13(24):4697–4710CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Bhatia SN, Ingber DE (2014) Microfluidic organs-on-chips. Nat Biotechnol 32(8):760–772CrossRefPubMedGoogle Scholar
  17. 17.
    Huh D, Torisawa YS, Hamilton GA et al (2012) Microengineered physiological biomimicry: organs-on-chips. Lab Chip 12(12):2156–2164CrossRefPubMedGoogle Scholar
  18. 18.
    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
  19. 19.
    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
  20. 20.
    Moya ML, Hsu YH, Lee AP et al (2013) In vitro perfused human capillary networks. Tissue Eng Part C Methods 19(9):730–737CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    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
  22. 22.
    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

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of Micro/Nano ElectronicsShanghai Jiao Tong UniversityShanghaiChina
  2. 2.Department of Molecular Biology and BiochemistryUniversity of CaliforniaIrvineUSA
  3. 3.Department of Biomedical EngineeringWashington University in St. LouisSaint LouisUSA
  4. 4.Department of Biomedical EngineeringUniversity of CaliforniaIrvineUSA
  5. 5.Edwards Lifesciences Center for Advanced Cardiovascular TechnologyUniversity of CaliforniaIrvineUSA
  6. 6.Department of Mechanical and Aerospace EngineeringUniversity of CaliforniaIrvineUSA

Personalised recommendations