Embryonic stem cell-derived astrocytes expressing drug-inducible transgenes: Differentiation and transplantion into the mouse brain

Ronald Benveniste, Gordon Keller, Isabelle Germano

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Object. Embryonic stem cell (ESC)-derived astrocytes have many theoretical and practical advantages as vectors for delivery of gene therapy to the central nervous system (CNS). The aim of this study was to generate highly pure populations of ESC-derived astrocytes expressing drug-inducible transgenes, while minimizing contamination by undifferentiated ESCs Methods. Embryonic stem cells carrying a doxycycline-inducible green fluorescent protein (GFP) transgene were induced to differentiate into astrocytes by using feeder cell-free conditions that are completely defined. More than 95% of these cells expressed the astrocyte markers glial fibrillary acidic protein and GLT-1 glutamate transporter, and the morphological characteristics of these cells were typical of astrocytes. The expression of additional astrocyte markers was detected using reverse transcription-polymerase chain reaction. Undifferentiated ESCs comprised fewer than 0.1% of the cells after 10 days in this culture. Positive and negative selection techniques based on fluorescence-activated cell sorting were successfully used to decrease further the numbers of undifferentiated ESCs. Fully differentiated astrocytes expressed a GFP transgene under the tight control of a doxycycline-responsive promoter, and maintained their astrocytic phenotype 24 hours after transplantation into the mouse brain. Conclusions. This study shows that transgenic ESCs can be induced to differentiate into highly pure populations of astrocytes. The astrocytes continue to express the transgene under the tight control of a drug-inducible promoter and are suitable for transplantation into the mouse brain. The number of potentially hazardous ESCs can be minimized using cell-sorting techniques. This strategy may be used to generate cellular vectors for delivering gene therapy to the CNS.

Original languageEnglish
Pages (from-to)115-123
Number of pages9
JournalJournal of Neurosurgery
Volume103
Issue number1
DOIs
StatePublished - Jul 1 2005
Externally publishedYes

Fingerprint

Embryonic Stem Cells
Transgenes
Astrocytes
Brain
Pharmaceutical Preparations
Doxycycline
Green Fluorescent Proteins
Genetic Therapy
Central Nervous System
Transplantation
Feeder Cells
Amino Acid Transport System X-AG
Drug and Narcotic Control
Glial Fibrillary Acidic Protein
Population
Reverse Transcription
Flow Cytometry
Phenotype
Polymerase Chain Reaction

Keywords

  • Astrocyte
  • Embryonic stem cell
  • Gene therapy
  • Teratoma
  • Transplantation

ASJC Scopus subject areas

  • Clinical Neurology
  • Neuroscience(all)

Cite this

Embryonic stem cell-derived astrocytes expressing drug-inducible transgenes : Differentiation and transplantion into the mouse brain. / Benveniste, Ronald; Keller, Gordon; Germano, Isabelle.

In: Journal of Neurosurgery, Vol. 103, No. 1, 01.07.2005, p. 115-123.

Research output: Contribution to journalArticle

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