Differentiation of Inflammation-Responsive Astrocytes from Glial Progenitors Generated from Human Induced Pluripotent Stem Cells

Renata Santos; Krishna C. Vadodaria; Baptiste N. Jaeger; Arianna Mei; Sabrina Lefcochilos-Fogelquist; Ana P.D. Mendes; Galina Erikson; Maxim Shokhirev; Lynne Randolph-Moore; Callie Fredlender; Sonia Dave; Ruth Oefner; Conor Fitzpatrick; Monique Pena; Jerika J. Barron; Manching Ku; Ahmet M. Denli; Bilal E. Kerman; Patrick Charnay; John R. Kelsoe; Maria C. Marchetto; Fred H. Gage

doi:10.1016/j.stemcr.2017.05.011

Stem Cell Reports (Jun 2017)

Differentiation of Inflammation-Responsive Astrocytes from Glial Progenitors Generated from Human Induced Pluripotent Stem Cells

Renata Santos,
Krishna C. Vadodaria,
Baptiste N. Jaeger,
Arianna Mei,
Sabrina Lefcochilos-Fogelquist,
Ana P.D. Mendes,
Galina Erikson,
Maxim Shokhirev,
Lynne Randolph-Moore,
Callie Fredlender,
Sonia Dave,
Ruth Oefner,
Conor Fitzpatrick,
Monique Pena,
Jerika J. Barron,
Manching Ku,
Ahmet M. Denli,
Bilal E. Kerman,
Patrick Charnay,
John R. Kelsoe,
Maria C. Marchetto,
Fred H. Gage

Affiliations

Renata Santos: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Krishna C. Vadodaria: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Baptiste N. Jaeger: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Arianna Mei: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Sabrina Lefcochilos-Fogelquist: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Ana P.D. Mendes: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Galina Erikson: The Razavi Newman Integrative Genomics and Bioinformatics Core Facility, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Maxim Shokhirev: The Razavi Newman Integrative Genomics and Bioinformatics Core Facility, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Lynne Randolph-Moore: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Callie Fredlender: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Sonia Dave: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Ruth Oefner: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Conor Fitzpatrick: Flow Cytometry Core Facility, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Monique Pena: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Jerika J. Barron: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Manching Ku: The Razavi Newman Integrative Genomics and Bioinformatics Core Facility, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Ahmet M. Denli: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Bilal E. Kerman: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Patrick Charnay: Ecole Normale Supérieure, PSL Research University, CNRS, Inserm, Institut de Biologie de l’Ecole Normale Supérieure (IBENS), 46 rue d’Ulm, 75005 Paris, France
John R. Kelsoe: Department of Psychiatry, VA San Diego Healthcare System, La Jolla, CA 92151, USA
Maria C. Marchetto: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
Fred H. Gage: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA

DOI: https://doi.org/10.1016/j.stemcr.2017.05.011
Journal volume & issue: Vol. 8, no. 6
pp. 1757 – 1769

Abstract

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Astrocyte dysfunction and neuroinflammation are detrimental features in multiple pathologies of the CNS. Therefore, the development of methods that produce functional human astrocytes represents an advance in the study of neurological diseases. Here we report an efficient method for inflammation-responsive astrocyte generation from induced pluripotent stem cells (iPSCs) and embryonic stem cells. This protocol uses an intermediate glial progenitor stage and generates functional astrocytes that show levels of glutamate uptake and calcium activation comparable with those observed in human primary astrocytes. Stimulation of stem cell-derived astrocytes with interleukin-1β or tumor necrosis factor α elicits a strong and rapid pro-inflammatory response. RNA-sequencing transcriptome profiling confirmed that similar gene expression changes occurred in iPSC-derived and primary astrocytes upon stimulation with interleukin-1β. This protocol represents an important tool for modeling in-a-dish neurological diseases with an inflammatory component, allowing for the investigation of the role of diseased astrocytes in neuronal degeneration.

Published in Stem Cell Reports

ISSN: 2213-6711 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Medicine: Medicine (General); Science: Biology (General)
Website: http://www.cell.com/stem-cell-reports/home

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