Advertisement

Organoids pp 101-112 | Cite as

Generation of Functional Kidney Organoids In Vivo Starting from a Single-Cell Suspension

  • Valentina Benedetti
  • Valerio Brizi
  • Christodoulos XinarisEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1576)

Abstract

Novel methods in developmental biology and stem cell research have made it possible to generate complex kidney tissues in vitro that resemble whole organs and are termed organoids. In this chapter we describe a technique using suspensions of fully dissociated mouse kidney cells to yield organoids that can become vascularized in vivo and mature and display physiological functions. This system can be used to produce fine-grained human–mouse chimeric organoids in which the renal differentiation potential of human cells can be assessed. It can also be an excellent method for growing chimeric organoids in vivo using human stem cells, which can differentiate into specialized kidney cells and exert nephron-specific functions. We provide detailed methods, a brief discussion of critical points, and describe some successfully implemented examples of the system.

Keywords:

Kidney organoids Stem cells Implantation Kidney engineering Glomerulogenesis Dissociation-reaggregation assay VEGF Cell suspensions Kidney development 

References

  1. 1.
    Auerbach R, Grobstein C (1958) Inductive interaction of embryonic tissues after dissociation and reaggregation. Exp Cell Res 15(2):384–397CrossRefGoogle Scholar
  2. 2.
    Unbekandt M, Davies JA (2010) Dissociation of embryonic kidneys followed by reaggregation allows the formation of renal tissues. Kidney Int 77(5):407–416CrossRefGoogle Scholar
  3. 3.
    Xinaris C, Benedetti V, Rizzo P, Abbate M, Corna D, Azzollini N, Conti S, Unbekandt M, Davies JA, Morigi M, Benigni A, Remuzzi G (2012) In vivo maturation of functional renal organoids formed from embryonic cell suspensions. J Am Soc Nephrol 23(11):1857–1868CrossRefGoogle Scholar
  4. 4.
    Xinaris C, Benedetti V, Novelli R, Abbate M, Rizzo P, Conti S, Tomasoni S, Corna D, Pozzobon M, Cavallotti D, Yokoo T, Morigi M, Benigni A, Remuzzi G (2015) Functional human podocytes generated in organoids from amniotic fluid stem cells. J Am Soc Nephrol 27(5):1400–1411CrossRefGoogle Scholar
  5. 5.
    Kitamoto Y, Tokunaga H, Tomita K (1997) Vascular endothelial growth factor is an essential molecule for mouse kidney development: glomerulogenesis and nephrogenesis. J Clin Invest 99(10):2351–2357CrossRefGoogle Scholar
  6. 6.
    Siegel N, Rosner M, Unbekandt M, Fuchs C, Slabina N, Dolznig H, Davies JA, Lubec G, Hengstschlager M (2010) Contribution of human amniotic fluid stem cells to renal tissue formation depends on mTOR. Hum Mol Genet 19(17):3320–3331CrossRefGoogle Scholar
  7. 7.
    Hendry CE, Vanslambrouck JM, Ineson J, Suhaimi N, Takasato M, Rae F, Little MH (2013) Direct transcriptional reprogramming of adult cells to embryonic nephron progenitors. J Am Soc Nephrol 24(9):1424–1434CrossRefGoogle Scholar
  8. 8.
    Papadimou E, Morigi M, Iatropoulos P, Xinaris C, Tomasoni S, Benedetti V, Longaretti L, Rota C, Todeschini M, Rizzo P, Introna M, Grazia de Simoni M, Remuzzi G, Goligorsky MS, Benigni A (2015) Direct reprogramming of human bone marrow stromal cells into functional renal cells using cell-free extracts. Stem Cell Reports 4(4):685–698CrossRefGoogle Scholar
  9. 9.
    Mae S, Shono A, Shiota F, Yasuno T, Kajiwara M, Gotoda-Nishimura N, Arai S, Sato-Otubo A, Toyoda T, Takahashi K, Nakayama N, Cowan CA, Aoi T, Ogawa S, McMahon AP, Yamanaka S, Osafune K (2013) Monitoring and robust induction of nephrogenic intermediate mesoderm from human pluripotent stem cells. Nat Commun 4:1367CrossRefGoogle Scholar
  10. 10.
    Xia Y, Nivet E, Sancho-Martinez I, Gallegos T, Suzuki K, Okamura D, Wu MZ, Dubova I, Esteban CR, Montserrat N, Campistol JM, Izpisua Belmonte JC (2013) Directed differentiation of human pluripotent cells to ureteric bud kidney progenitor-like cells. Nat Cell Biol 15(12):1507–1515CrossRefGoogle Scholar
  11. 11.
    Takasato M, Er PX, Becroft M, Vanslambrouck JM, Stanley EG, Elefanty AG, Little MH (2014) Directing human embryonic stem cell differentiation towards a renal lineage generates a self-organizing kidney. Nat Cell Biol 16(1):118–126CrossRefGoogle Scholar
  12. 12.
    Taguchi A, Kaku Y, Ohmori T, Sharmin S, Ogawa M, Sasaki H, Nishinakamura R (2014) Redefining the in vivo origin of metanephric nephron progenitors enables generation of complex kidney structures from pluripotent stem cells. Cell Stem Cell 14(1):53–67CrossRefGoogle Scholar
  13. 13.
    Davies JA (2010) The embryonic kidney: isolation, organ culture, immunostaining and RNA interference. Methods Mol Biol 633:57–69CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 2.5 International License (http://creativecommons.org/licenses/by-nc/2.5/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

The images or other third party material in this chapter are included in the chapter's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the chapter's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

Authors and Affiliations

  • Valentina Benedetti
    • 1
  • Valerio Brizi
    • 1
  • Christodoulos Xinaris
    • 1
    Email author
  1. 1.IRCCS-Istituto di Ricerche Farmacologiche ‘Mario Negri’, Centro Anna Maria AstoriBergamoItaly

Personalised recommendations