Alzheimer’s disease is the most common form of dementia, characterized by two pathological hallmarks: amyloid-β plaques and neurofibrillary tangles1. The amyloid hypothesis of Alzheimer’s disease posits that the excessive accumulation of amyloid-β peptide leads to neurofibrillary tangles composed of aggregated hyperphosphorylated tau2,3. However, to date, no single disease model has serially linked these two pathological events using human neuronal cells. Mouse models with familial Alzheimer’s disease (FAD) mutations exhibit amyloid-β-induced synaptic and memory deficits but they do not fully recapitulate other key pathological events of Alzheimer’s disease, including distinct neurofibrillary tangle pathology4,5. Human neurons derived from Alzheimer’s disease patients have shown elevated levels of toxic amyloid-β species and phosphorylated tau but did not demonstrate amyloid-β plaques or neurofibrillary tangles6,7,8,9,10,11. Here we report that FAD mutations in β-amyloid precursor protein and presenilin 1 are able to induce robust extracellular deposition of amyloid-β, including amyloid-β plaques, in a human neural stem-cell-derived three-dimensional (3D) culture system. More importantly, the 3D-differentiated neuronal cells expressing FAD mutations exhibited high levels of detergent-resistant, silver-positive aggregates of phosphorylated tau in the soma and neurites, as well as filamentous tau, as detected by immunoelectron microscopy. Inhibition of amyloid-β generation with β- or γ-secretase inhibitors not only decreased amyloid-β pathology, but also attenuated tauopathy. We also found that glycogen synthase kinase 3 (GSK3) regulated amyloid-β-mediated tau phosphorylation. We have successfully recapitulated amyloid-β and tau pathology in a single 3D human neural cell culture system. Our unique strategy for recapitulating Alzheimer’s disease pathology in a 3D neural cell culture model should also serve to facilitate the development of more precise human neural cell models of other neurodegenerative disorders.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from $8.99

All prices are NET prices.


  1. 1.

    & Twenty years of the Alzheimer’s disease amyloid hypothesis: a genetic perspective. Cell 120, 545–555 (2005)

  2. 2.

    & The amyloid hypothesis of Alzheimer’s disease: progress and problems on the road to therapeutics. Science 297, 353–356 (2002)

  3. 3.

    Alzheimer’s disease is a synaptic failure. Science 298, 789–791 (2002)

  4. 4.

    Transgenic mouse models of Alzheimer’s disease: phenotype and mechanisms of pathogenesis. Biochem. Soc. Symp. 67, 195–202 (2001)

  5. 5.

    Selecting a mouse model of Alzheimer’s disease. Methods Mol. Biol. 670, 169–189 (2011)

  6. 6.

    & iPSCs to the rescue in Alzheimer’s research. Cell Stem Cell 10, 235–236 (2012)

  7. 7.

    et al. Modeling familial Alzheimer’s disease with induced pluripotent stem cells. Hum. Mol. Genet. 20, 4530–4539 (2011)

  8. 8.

    et al. Probing sporadic and familial Alzheimer’s disease using induced pluripotent stem cells. Nature 482, 216–220 (2012)

  9. 9.

    et al. Modeling Alzheimer’s disease with iPSCs reveals stress phenotypes associated with intracellular Aβ and differential drug responsiveness. Cell Stem Cell 12, 487–496 (2013)

  10. 10.

    et al. The familial Alzheimer’s disease APPV717I mutation alters APP processing and Tau expression in iPSC-derived neurons. Hum. Mol. Genet. 23, 3523–3536 (2014)

  11. 11.

    et al. Characterization and molecular profiling of PSEN1 familial Alzheimer’s disease iPSC-derived neural progenitors. PLoS ONE 9, e84547 (2014)

  12. 12.

    et al. Differential development of neuronal physiological responsiveness in two human neural stem cell lines. BMC Neurosci. 8, 36 (2007)

  13. 13.

    et al. Protection of neurons derived from human neural progenitor cells by veratridine. Neuroreport 20, 1225–1229 (2009)

  14. 14.

    et al. Quantitative and kinetic profile of Wnt/β-catenin signaling components during human neural progenitor cell differentiation. Cell. Mol. Biol. Lett. 16, 515–538 (2011)

  15. 15.

    , & 3D differentiation of neural stem cells in macroporous photopolymerizable hydrogel scaffolds. PLoS ONE 7, e48824 (2012)

  16. 16.

    et al. Cerebral organoids model human brain development and microcephaly. Nature 501, 373–379 (2013)

  17. 17.

    et al. Bioengineered functional brain-like cortical tissue. Proc. Natl Acad. Sci. USA (August 11, 2014)

  18. 18.

    , & Tau protein and tauopathy. Neuropsychopharmacology: The Fifth Generation of Progress (eds , , & ) 1339–1354 (Lippincott, Williams & Wilkins, 2002)

  19. 19.

    et al. Soluble γ-secretase modulators selectively inhibit the production of the 42-amino acid amyloid β peptide variant and augment the production of multiple carboxy-truncated amyloid β species. Biochemistry 53, 702–713 (2014)

  20. 20.

    & The role of tau in Alzheimer’s disease. Med. Clin. North Am. 86, 615–627 (2002)

  21. 21.

    et al. Neuronal and microglial involvement in β-amyloid protein deposition in Alzheimer’s disease. Am. J. Pathol. 137, 241–246 (1990)

  22. 22.

    & Use of excitatory amino acids to make axon-sparing lesions of hypothalamus. Methods Enzymol. 103, 393–400 (1983)

  23. 23.

    , , , & 1-Azakenpaullone is a selective inhibitor of glycogen synthase kinase-3β. Bioorg. Med. Chem. Lett. 14, 413–416 (2004)

  24. 24.

    et al. Glycogen synthase kinase 3 (GSK3) inhibitor, SB-216763, promotes pluripotency in mouse embryonic stem cells. PLoS ONE 7, e39329 (2012)

  25. 25.

    , , & GSK-3α regulates production of Alzheimer’s disease amyloid-β peptides. Nature 423, 435–439 (2003)

  26. 26.

    et al. GSK-3α/β kinases and amyloid production in vivo. Nature 423, 435–439 (2003)

  27. 27.

    et al. A transgenic Alzheimer rat with plaques, tau pathology, behavioral impairment, oligomeric Aβ, and frank neuronal loss. J. Neurosci. 33, 6245–6256 (2013)

  28. 28.

    et al. Acceleration and persistence of neurofibrillary pathology in a mouse model of tauopathy following anesthesia. FASEB J. 23, 2595–2604 (2009)

  29. 29.

    , & Biochemical isolation of insoluble tau in transgenic mouse models of tauopathies. Methods Mol. Biol. 849, 473–491 (2012)

  30. 30.

    , , , & Human stem cell-derived neurons: a system to study human tau function and dysfunction. PLoS ONE 5, e13947 (2010)

Download references


This work was supported by the grants from the Cure Alzheimer’s fund (D.Y.K., S.H.C. and R.E.T.) and national Institute of Health grants 5P01AG15379 (R.E.T.) and 5R37MH060009 (R.E.T.). We thank T. L. Spires, M. Polydoro and S. Wegmann for revising the manuscript, and M. L. McKee for the electron microscopy assistance. We also appreciate B. T. Hyman, O. Berezovska, J. Hardy and P. Davies for providing cDNAs and antibodies. We acknowledge Ragon Institute’s Imaging Core facility (part of the Harvard CFAR Immunology Core), Massachusetts General Hospital (MGH) Viral Vector Core (supported by NIH/NINDS P30NS04776), MGH Microscopy Core of the Center for Systems Biology for immunoelectron microscopy (partially supported by an IBDG Grant DK43351 and a BADERC Award DK57521), MGH Confocal Microscope Core and MGH Pathology Core for technical and instrument support.

Author information

Author notes

    • Se Hoon Choi
    •  & Young Hye Kim

    These authors contributed equally to this work.


  1. Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, USA

    • Se Hoon Choi
    • , Young Hye Kim
    • , Matthias Hebisch
    • , Christopher Sliwinski
    • , Carla D’Avanzo
    • , Hechao Chen
    • , Basavaraj Hooli
    • , Caroline Asselin
    • , Justin B. Klee
    • , Can Zhang
    • , Dora M. Kovacs
    • , Rudolph E. Tanzi
    •  & Doo Yeon Kim
  2. Division of Mass Spectrometry Research, Korea Basic Science Institute, Cheongju-si, Chungbuk 363-883, South Korea

    • Young Hye Kim
  3. Institute of Reconstructive Neurobiology, Life and Brain Center, University of Bonn and Hertie Foundation, 53127 Bonn, Germany

    • Matthias Hebisch
    •  & Michael Peitz
  4. FM Kirby Neurobiology Center, Boston Children’s Hospital and Harvard Stem Cell Institute, Boston, Massachusetts 02115, USA

    • Seungkyu Lee
    • , Brian J. Wainger
    •  & Clifford J. Woolf
  5. The Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA

    • Julien Muffat
  6. Department of Neurosciences, University of California, San Diego, La Jolla, California 92093, USA

    • Steven L. Wagner


  1. Search for Se Hoon Choi in:

  2. Search for Young Hye Kim in:

  3. Search for Matthias Hebisch in:

  4. Search for Christopher Sliwinski in:

  5. Search for Seungkyu Lee in:

  6. Search for Carla D’Avanzo in:

  7. Search for Hechao Chen in:

  8. Search for Basavaraj Hooli in:

  9. Search for Caroline Asselin in:

  10. Search for Julien Muffat in:

  11. Search for Justin B. Klee in:

  12. Search for Can Zhang in:

  13. Search for Brian J. Wainger in:

  14. Search for Michael Peitz in:

  15. Search for Dora M. Kovacs in:

  16. Search for Clifford J. Woolf in:

  17. Search for Steven L. Wagner in:

  18. Search for Rudolph E. Tanzi in:

  19. Search for Doo Yeon Kim in:


D.Y.K. and R.E.T. were equally responsible for experimental design and data interpretation. S.H.C., Y.H.K. and D.Y.K. mainly contributed to writing and revising the manuscript. D.Y.K., Y.H.K., S.H.C., M.H., S.L., C.D., H.C., C.S., B.H., J.B.K., C.A. and C.Z. conducted the experiments. S.L.W. synthesized SGSM41 and S.L.W. and C.Z. characterized SGSM41. J.M., B.J.W., M.P., C.J.W. and D.M.K. contributed to data interpretation.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Rudolph E. Tanzi or Doo Yeon Kim.

Extended data

Supplementary information


  1. 1.

    3D reconstitution of ReN-G cells after 2-week differentiation in 3D Matrigel (green, GFP; 400x magnification)

    Due to the technical limitation of spinning disk confocal microscopy (Olympus), 3D reconstituted images represent only a part of the whole 3D culture.

  2. 2.

    3D reconstitution of ReN-m (enriched) cells after 4-week differentiation in 3D Matrigel (red, mCherry; 400x magnification)

    3D reconstitution of ReN-m (enriched) cells after 4-week differentiation in 3D Matrigel (red, mCherry; 400x magnification)

  3. 3.

    3D reconstitution of 3D-differentiated ReN-m cells for 6 weeks (green, Synapsin I; red, mCherry; 1,000x magnification).

    3D reconstitution of 3D-differentiated ReN-m cells for 6 weeks (green, Synapsin I; red, mCherry; 1,000x magnification).

  4. 4.

    Z-section sequences of 3D6-stained Aβ aggregates in 6-week differentiated ReN-mGAP cells (green, GFP; 400x magnification).

    Z-section sequences of 3D6-stained Aβ aggregates in 6-week differentiated ReN-mGAP cells (green, GFP; 400x magnification).

About this article

Publication history







By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.