Direct Cell Reprogramming of Mouse Fibroblasts into Functional Astrocytes Using Lentiviral Overexpression of the Transcription Factors NFIA, NFIB, and SOX9

Boning Qiu, Ruben J de Vries, Massimiliano Caiazzo*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Abstract

Astrocytes play an important role in maintaining brain homeostasis and their dysfunction is involved in a number of neurological disorders. An accessible source of astrocytes is essential to model neurological diseases and potential cell therapy approaches. Cell reprogramming techniques offer possibilities to reprogram terminally differentiated cells into other cell types. By overexpressing the three astrocytic transcription factors NFIA, NFIB, and SOX9, we showed that it is possible to directly transdifferentiate fibroblasts into functional astrocytes. These induced astrocytes (iAstrocytes) express glial fibrillary acidic protein (GFAP) and S100 calcium binding protein B (S100B), as well as other astrocytic markers. Moreover, electrophysiological properties indicate that iAstrocytes are functionally comparable to native brain astrocytes. Here we describe an optimized protocol to generate iAstrocytes starting from skin fibroblasts and this approach can be adapted for a wide range of somatic cell types.

Original languageEnglish
Title of host publicationNeural Reprogramming
Subtitle of host publicationMethods and Protocols
EditorsHenrik Ahlenius
PublisherSpringer
Chapter3
Pages31-43
Number of pages13
Volume2352
ISBN (Electronic)978-1-0716-1601-7
ISBN (Print)978-1-0716-1600-0, Softcover ISBN 978-1-0716-1603-1
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2352
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Astrocytes
  • Cell reprogramming
  • Fibroblasts
  • Lentivirus
  • Transcription factors

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