This article has been updated


Our understanding of Alzheimer’s disease pathogenesis is currently limited by difficulties in obtaining live neurons from patients and the inability to model the sporadic form of the disease. It may be possible to overcome these challenges by reprogramming primary cells from patients into induced pluripotent stem cells (iPSCs). Here we reprogrammed primary fibroblasts from two patients with familial Alzheimer’s disease, both caused by a duplication of the amyloid-β precursor protein gene1 (APP; termed APPDp), two with sporadic Alzheimer’s disease (termed sAD1, sAD2) and two non-demented control individuals into iPSC lines. Neurons from differentiated cultures were purified with fluorescence-activated cell sorting and characterized. Purified cultures contained more than 90% neurons, clustered with fetal brain messenger RNA samples by microarray criteria, and could form functional synaptic contacts. Virtually all cells exhibited normal electrophysiological activity. Relative to controls, iPSC-derived, purified neurons from the two APPDp patients and patient sAD2 exhibited significantly higher levels of the pathological markers amyloid-β(1–40), phospho-tau(Thr 231) and active glycogen synthase kinase-3β (aGSK-3β). Neurons from APPDp and sAD2 patients also accumulated large RAB5-positive early endosomes compared to controls. Treatment of purified neurons with β-secretase inhibitors, but not γ-secretase inhibitors, caused significant reductions in phospho-Tau(Thr 231) and aGSK-3β levels. These results suggest a direct relationship between APP proteolytic processing, but not amyloid-β, in GSK-3β activation and tau phosphorylation in human neurons. Additionally, we observed that neurons with the genome of one sAD patient exhibited the phenotypes seen in familial Alzheimer’s disease samples. More generally, we demonstrate that iPSC technology can be used to observe phenotypes relevant to Alzheimer’s disease, even though it can take decades for overt disease to manifest in patients.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Change history

  • 08 February 2012

    An addition was made to Acknowlegements.


Primary accessions

Gene Expression Omnibus

Data deposits

Data have been deposited in the Gene ExpressionOmnibus under accession GSE34879.


  1. 1.

    et al. APP locus duplication in a Finnish family with dementia and intracerebral haemorrhage. J. Neurol. Neurosurg. Psychiatry 78, 1158–1159 (2007)

  2. 2.

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

  3. 3.

    , & Tau-mediated neurodegeneration in Alzheimer’s disease and related disorders. Nature Rev. Neurosci. 8, 663–672 (2007)

  4. 4.

    et al. Role of genes and environments for explaining Alzheimer disease. Arch. Gen. Psychiatry 63, 168–174 (2006)

  5. 5.

    et al. Alzheimer-type neuropathology in transgenic mice overexpressing V717F β-amyloid precursor protein. Nature 373, 523–527 (1995)

  6. 6.

    et al. Reducing endogenous tau ameliorates amyloid β-induced deficits in an Alzheimer’s disease mouse model. Science 316, 750–754 (2007)

  7. 7.

    , , & β-Amyloid fibrils induce tau phosphorylation and loss of microtubule binding. Neuron 14, 879–888 (1995)

  8. 8.

    et al. Induced pluripotent stem cells from a spinal muscular atrophy patient. Nature 457, 277–280 (2009)

  9. 9.

    et al. LRRK2 mutant iPSC-derived DA neurons demonstrate increased susceptibility to oxidative stress. Cell Stem Cell 8, 267–280 (2011)

  10. 10.

    et al. Directed conversion of Alzheimer’s disease patient skin fibroblasts into functional neurons. Cell 3, 359–371 (2011)

  11. 11.

    , , & Differential modeling of fragile X syndrome by human embryonic stem cells and induced pluripotent stem cells. Cell Stem Cell 6, 407–411 (2010)

  12. 12.

    et al. Cell-surface marker signatures for the isolation of neural stem cells, glia and neurons derived from human pluripotent stem cells. PLoS ONE 6, e17540 (2011)

  13. 13.

    , & YAP regulates neural progenitor cell number via the TEA domain transcription factor. Genes Dev. 22, 3320–3334 (2008)

  14. 14.

    & Regulation of Drosophila glial cell proliferation by Merlin-Hippo signaling. Development 138, 5201–5212 (2011)

  15. 15.

    et al. Excessive production of amyloid β-protein by peripheral cells of symptomatic and presymptomatic patients carrying the Swedish familial Alzheimer disease mutation. Proc. Natl Acad. Sci. USA 91, 11993–11997 (1994)

  16. 16.

    et al. Secreted amyloid β-protein similar to that in the senile plaques of Alzheimer’s disease is increased in vivo by the presenilin 1 and 2 and APP mutations linked to familial Alzheimer’s disease. Nature Med. 2, 864–870 (1996)

  17. 17.

    et al. Peripheral markers in testing pathophysiological hypotheses and diagnosing Alzheimer’s disease. FASEB J. 12, 17–34 (1998)

  18. 18.

    , , & Neurofibrillary tangles but not senile plaques parallel duration and severity of Alzheimer’s disease. Neurology 42, 631 (1992)

  19. 19.

    & Primed phosphorylation of tau at Thr 231 by glycogen synthase kinase 3β (GSK3β) plays a critical role in regulating tau’s ability to bind and stabilize microtubules. J. Neurochem. 88, 349–358 (2004)

  20. 20.

    et al. CSF tau protein phosphorylated at threonine 231 correlates with cognitive decline in MCI subjects. Neurology 59, 627–629 (2002)

  21. 21.

    et al. CSF phosphorylated tau protein correlates with neocortical neurofibrillary pathology in Alzheimer’s disease. Brain 129, 3035–3041 (2006)

  22. 22.

    & Glycogen synthase kinase 3β phosphorylates tau at both primed and unprimed sites. J. Biol. Chem. 278, 187–193 (2003)

  23. 23.

    et al. Crystal structure of glycogen synthase kinase 3β: structural basis for phosphate-primed substrate specificity and autoinhibition. Cell 105, 721–732 (2001)

  24. 24.

    et al. Endocytic pathway abnormalities precede amyloid β deposition in sporadic Alzheimer’s disease and Down syndrome: differential effects of APOE genotype and presenilin mutations. Am. J. Pathol. 157, 277–286 (2000)

  25. 25.

    et al. Endocytic disturbances distinguish among subtypes of Alzheimer’s disease and related disorders. Ann. Neurol. 50, 661–665 (2001)

  26. 26.

    Endosome function and dysfunction in Alzheimer’s disease and other neurodegenerative diseases. Neurobiol. Aging 26, 373–382 (2005)

  27. 27.

    , & Synaptic loss in Alzheimer’s disease and other dementias. Neurology 39, 355–361 (1989)

  28. 28.

    , , & Regional alteration of synapsin I in the hippocampal formation of Alzheimer’s disease patients. Acta Neuropathol. 107, 209–215 (2004)

  29. 29.

    et al. Increased App expression in a mouse model of Down’s syndrome disrupts NGF transport and causes cholinergic neuron degeneration. Neuron 51, 29–42 (2006)

  30. 30.

    et al. Alzheimer’s-related endosome dysfunction in Down syndrome is Aβ-independent but requires APP and is reversed by BACE-1 inhibition. Proc. Natl Acad. Sci. USA 107, 1630–1635 (2010)

  31. 31.

    in Current Protocols in Cell Biology Ch. 2 12 (John Wiley & Sons, 2001)

  32. 32.

    , , , & Generation of human-induced pluripotent stem cells. Nature Protocols 3, 1180–1186 (2008)

  33. 33.

    , & Refractory nature of normal human diploid fibroblasts with respect to oncogene-mediated transformation. Proc. Natl Acad. Sci. USA 100, 13567–13572 (2003)

  34. 34.

    et al. The ROCK inhibitor Y-27632 improves recovery of human embryonic stem cells after fluorescence-activated cell sorting with multiple cell surface markers. PLoS ONE 5, e12148 (2010)

  35. 35.

    et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872 (2007)

  36. 36.

    et al. Induction of midbrain dopaminergic neurons from ES cells by stromal cell derived inducing activity. Neuron 28, 31–40 (2000)

Download references


We thank D. Galasko, M. Sundsmo, J. Rivera, J. Fontaine, C. Gigliotti and B. Yu at the University of California, San Diego (UCSD) Alzheimer’s Disease Research Center for patient samples and data (grant AGO 5131); S. Dowdy and N. Yoshioka for viral vectors; B. Balderas at BD Biosciences for antibodies; C. Santucci and S. Nguyen for teratoma assay assistance; the UCSD Neuroscience Microscopy Shared Facility (grant P30 NS047101); and Planned Parenthood of the Pacific Southwest for fetal brain specimens. Funding was from California Institute of Regenerative Medicine (CIRM) comprehensive grants (M.M., F.H.G., L.S.B.G.), CIRM predoctoral fellowship (M.A.I.), FP7 Marie Curie IOF (C.B.), Weatherstone Foundation fellowship (K.L.N.), National Institutes of Health K12 HD001259, the Hartwell Foundation (L.C.L., F.S.B.), the Lookout Fund and the McDonnell Foundation (F.H.G.). L.S.B.G. is an investigator with the Howard Hughes Medical Institute. Some experiments were conducted in J.F. Loring's laboratory (The Scripps Research Institute) with support from grants TR1-01250, CL1-00502, RM1-01717 (CIRM) and a gift from the Esther O'Keefe Foundation.

Author information


  1. Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA

    • Mason A. Israel
    • , Shauna H. Yuan
    • , Sol M. Reyna
    • , Cheryl Herrera
    •  & Lawrence S. B. Goldstein
  2. Biomedical Sciences Graduate Program, University of California, San Diego, La Jolla, California 92093, USA

    • Mason A. Israel
    •  & Sol M. Reyna
  3. Department of Neurosciences, University of California, San Diego, La Jolla, California, USA

    • Shauna H. Yuan
    • , Edward H. Koo
    •  & Lawrence S. B. Goldstein
  4. The Salk Institute for Biological Studies, La Jolla, California 92037, USA

    • Cedric Bardy
    • , Yangling Mu
    •  & Fred H. Gage
  5. Department of Anesthesiology, University of California, San Diego, La Jolla, California 92093, USA

    • Michael P. Hefferan
    •  & Martin Marsala
  6. Department of Anesthesiology, Maastricht University Medical Center, Maastricht 6202 AZ, Netherlands

    • Sebastiaan Van Gorp
  7. Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA

    • Kristopher L. Nazor
  8. Department of Reproductive Medicine, University of California, San Diego, La Jolla, California 92093, USA

    • Francesca S. Boscolo
    •  & Louise C. Laurent
  9. BD Biosciences, La Jolla, California 92037, USA

    • Christian T. Carson
  10. Institute of Neurobiology, Slovak Academy of Sciences, Kosice SK-04001, Slovakia

    • Martin Marsala
  11. Department of Clinical Medicine, Neurology and Clinical Research Center, University of Oulu, Oulu FIN-90015, Finland

    • Anne M. Remes


  1. Search for Mason A. Israel in:

  2. Search for Shauna H. Yuan in:

  3. Search for Cedric Bardy in:

  4. Search for Sol M. Reyna in:

  5. Search for Yangling Mu in:

  6. Search for Cheryl Herrera in:

  7. Search for Michael P. Hefferan in:

  8. Search for Sebastiaan Van Gorp in:

  9. Search for Kristopher L. Nazor in:

  10. Search for Francesca S. Boscolo in:

  11. Search for Christian T. Carson in:

  12. Search for Louise C. Laurent in:

  13. Search for Martin Marsala in:

  14. Search for Fred H. Gage in:

  15. Search for Anne M. Remes in:

  16. Search for Edward H. Koo in:

  17. Search for Lawrence S. B. Goldstein in:


M.A.I. and L.S.B.G. conceived the project; M.A.I. and L.S.B.G. designed the experiments; M.A.I., S.H.Y., C.B., S.M.R., Y.M., C.H., M.P.H., S.V.G., M.M., K.L.N. and F.S.B. performed the experiments; M.A.I., S.H.Y. and C.T.C. developed differentiation methods; A.M.R. and E.H.K. provided APPDp patient samples and information; F.H.G. supervised C.B. and Y.M.; M.M. supervised M.P.H. and S.V.G.; L.C.L. supervised K.L.N. and F.S.B.; M.A.I. and L.S.B.G. wrote the manuscript; F.H.G., E.H.K. and A.M.R. edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Lawrence S. B. Goldstein.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-12 with legends and Supplementary Tables 1, 3 and 4 (see separate files for Supplementary Tables 2 and 5).

  2. 2.

    Supplementary Table 5

    This table contains Amyloid-β, aGSK3β and p-tau/total tau detailed by patient, iPSC line and culture well.

Excel files

  1. 1.

    Supplementary Table 2

    This table shows differentially expressed genes between purified neurons and fetal brain versus non-neural fetal samples (sheet 1) and differentially expressed genes between purified neurons versus fetal brain (sheet 2).

About this article

Publication history





Further reading


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.