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Endothelial Cells: Co-culture Spheroids

  • Janos M. Kanczler
  • Julia A. Wells
  • Richard O. C. OreffoEmail author
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 2206)

Abstract

the development and maintenance of a functioning vascular system is a critical function for many aspects of tissue growth and regeneration. vascular endothelial cell in vitro co-culture spheroids are self-organized cell composites that have the capacity to recapitulate the three-dimensional tissue microenvironment. these spheroid testing platforms aim to better understand the mechanisms of functional tissue and how new therapeutic agents can drive these 3d co-culture processes. here we describe direct cell–cell 3d endothelial co-culture spheroid methods, to examine the physiological spatial growth and cell–cell interaction of vascular cells and surrounding native tissue cells in the formation of vascular networks within spheroids and the potential to regenerate tissue.

Key words

Endothelial cells Co-culture Human bone marrow stromal cells Spheroids 3D Tissue engineering 

Notes

Acknowledgements

funding to roco from the biotechnology and biological sciences research council uk (bbsrc lo21071/ and bb/l00609x/1), the uk regenerative medicine platform acellular/smart materials—3d architecture (mr/r015651/1), and a grant from the uk regenerative medicine platform (mr/l012626/1 southampton imaging) is gratefully acknowledged.

References

  1. 1.
    Lilly B (2014) We have contact: endothelial cell-smooth muscle cell interactions. Physiology 29(4):234–241
  2. 2.
    Sweeney M, Foldes G (2018) It takes two: endothelial-perivascular cell cross-talk in vascular development and disease. Front Cardiovasc Med 5:154
  3. 3.
    Lazzari G, Nicolas V, Matsusaki M, Akashi M, Couvreur P, Mura S (2018) Multicellular spheroid based on a triple co-culture: a novel 3D model to mimic pancreatic tumor complexity. Acta Biomater 78:296–307
  4. 4.
    Shoval H, Karsch-Bluman A, Brill-Karniely Y, Stern T, Zamir G, Hubert A, Benny O (2017) Tumor cells and their crosstalk with endothelial cells in 3D spheroids. Sci Rep 7(1):10428
  5. 5.
    Guerrero J, Oliveira H, Catros S, Siadous R, Derkaoui S-M, Bareille R, Letourneur D, Amédée J (2014) The use of total human bone marrow fraction in a direct three-dimensional expansion approach for bone tissue engineering applications: focus on angiogenesis and osteogenesis. Tissue Eng A 21(5–6):861–874
  6. 6.
    Inglis S, Kanczler JM, Oreffo RO (2018) 3D human bone marrow stromal and endothelial cell spheres promote bone healing in an osteogenic niche. FASEB J 33(3):3279–3290
  7. 7.
    Walser R, Metzger W, Görg A, Pohlemann T, Menger M, Laschke M (2013) Generation of co-culture spheroids as vascularisation units for bone tissue engineering. Eur Cell Mater 26:222–233
  8. 8.
    Korff T, Augustin HG (1998) Integration of endothelial cells in multicellular spheroids prevents apoptosis and induces differentiation. J Cell Biol 143(5):1341–1352
  9. 9.
    Pfisterer L, Korff T (2016) Spheroid-based in vitro angiogenesis model. In: Angiogenesis protocols, 3rd edn. Springer, New York, NY, pp 167–177
  10. 10.
    Chiew GGY, Wei N, Sultania S, Lim S, Luo KQ (2017) Bioengineered three-dimensional co-culture of cancer cells and endothelial cells: a model system for dual analysis of tumor growth and angiogenesis. Biotechnol Bioeng 114(8):1865–1877
  11. 11.
    Carano RA, Filvaroff EH (2003) Angiogenesis and bone repair. Drug Discov Today 8(21):980–989
  12. 12.
    Kanczler J, Oreffo R (2008) Osteogenesis and angiogenesis: the potential for engineering bone. Eur Cell Mater 15(2):100–114
  13. 13.
    Bruder SP, Caplan AI (1989) Cellular and molecular events during embryonic bone development. Connect Tissue Res 20(1–4):65–71
  14. 14.
    Freeman FE, Haugh MG, McNamara LM (2015) An in vitro bone tissue regeneration strategy combining chondrogenic and vascular priming enhances the mineralization potential of mesenchymal stem cells in vitro while also allowing for vessel formation. Tissue Eng A 21(7–8):1320–1332
  15. 15.
    Shah S, Lee H, Park YH, Jeon E, Chung HK, Lee ES, Shim JH, Kang K-T (2019) Three-dimensional angiogenesis assay system using co-culture spheroids formed by endothelial colony forming cells and mesenchymal stem cells. J Vis Exp 151:e60032
  16. 16.
    Wimmer RA, Leopoldi A, Aichinger M, Kerjaschki D, Penninger JM (2019) Generation of blood vessel organoids from human pluripotent stem cells. Nat Protoc 14(11):3082–3100
  17. 17.
    Jaffe EA, Nachman RL, Becker CG, Minick CR (1973) Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 52(11):2745–2756
  18. 18.
    Oreffo RO, Driessens FC, Planell JA, Triffitt JT (1998) Growth and differentiation of human bone marrow osteoprogenitors on novel calcium phosphate cements. Biomaterials 19(20):1845–1854
  19. 19.
    Inglis S (2017) The role of the vasculature in skeletal development and repair – clues to improved regenerative strategies. Ph.D. Southampton, Southampton
  20. 20.
    Hoarau-Véchot J, Rafii A, Touboul C, Pasquier J (2018) Halfway between 2D and animal models: are 3D cultures the ideal tool to study cancer-microenvironment interactions? Int J Mol Sci 19(1):181

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • Janos M. Kanczler
    • 1
  • Julia A. Wells
    • 1
  • Richard O. C. Oreffo
    • 1
    Email author
  1. 1.Bone and Joint Research Group, Centre for Human Development, Stem Cells and Regeneration, Faculty of MedicineUniversity of SouthamptonSouthamptonUK

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