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Proc Natl Acad Sci U S A. 2018 Jun 19;115(25):6470-6475. doi: 10.1073/pnas.1720273115. Epub 2018 Jun 4.

Transdifferentiation of human adult peripheral blood T cells into neurons.

Author information

1
Department of Pathology, Stanford University, Stanford, CA 94305.
2
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA 94305.
3
Department of Bioengineering, Stanford University, Stanford, CA 94305.
4
Department of Molecular and Cellular Physiology and Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305.
5
Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305.
6
Department of Molecular and Cellular Physiology and Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305; tcs1@stanford.edu wernig@stanford.edu.
7
Department of Pathology, Stanford University, Stanford, CA 94305; tcs1@stanford.edu wernig@stanford.edu.

Abstract

Human cell models for disease based on induced pluripotent stem (iPS) cells have proven to be powerful new assets for investigating disease mechanisms. New insights have been obtained studying single mutations using isogenic controls generated by gene targeting. Modeling complex, multigenetic traits using patient-derived iPS cells is much more challenging due to line-to-line variability and technical limitations of scaling to dozens or more patients. Induced neuronal (iN) cells reprogrammed directly from dermal fibroblasts or urinary epithelia could be obtained from many donors, but such donor cells are heterogeneous, show interindividual variability, and must be extensively expanded, which can introduce random mutations. Moreover, derivation of dermal fibroblasts requires invasive biopsies. Here we show that human adult peripheral blood mononuclear cells, as well as defined purified T lymphocytes, can be directly converted into fully functional iN cells, demonstrating that terminally differentiated human cells can be efficiently transdifferentiated into a distantly related lineage. T cell-derived iN cells, generated by nonintegrating gene delivery, showed stereotypical neuronal morphologies and expressed multiple pan-neuronal markers, fired action potentials, and were able to form functional synapses. These cells were stable in the absence of exogenous reprogramming factors. Small molecule addition and optimized culture systems have yielded conversion efficiencies of up to 6.2%, resulting in the generation of >50,000 iN cells from 1 mL of peripheral blood in a single step without the need for initial expansion. Thus, our method allows the generation of sufficient neurons for experimental interrogation from a defined, homogeneous, and readily accessible donor cell population.

KEYWORDS:

direct conversion; disease modeling; iN cells; induced neuronal cells; transdifferentiation

PMID:
29866841
PMCID:
PMC6016798
DOI:
10.1073/pnas.1720273115
[Indexed for MEDLINE]
Free PMC Article

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