Human schwann cells retain essential phenotype characteristics after immortalization

Helmar C. Lehmann, Weiran Chen, Ruifa Mi, Shuo Wang, Ying Liu, Mahendra Rao, Ahmet Hoke

Research output: Contribution to journalArticle

Abstract

Schwann cells (SCs) play an important role in the pathogenesis of peripheral nerve diseases and represent a potential target for development of therapies. However, use of primary human SCs (hSCs) for in vitro models is limited because these cells are difficult to prepare and maintain in high yield and purity under common cell culture conditions. To circumvent this obstacle, we immortalized primary human fetal SCs using the SV40 large T-antigen and human telomerase reverse transcriptase expression vectors. After cloning, selection, and purification, we evaluated several immortalized SC lines for their ability to express extracellular matrix (ECM) molecules and myelinate embryonic rat sensory axons. In addition, we established a gene expression profile and explored their sensitivity to oxidative stress in a simple in vitro assay. Immortalized hSC clones expressed common glial markers and a broad variety of growth factors, receptors, and ECM molecules as determined by immunocytochemistry, microarray, and quantitative reverse transcription- polymerase chain reaction. In neuron-SC co-cultures, these cells were able to myelinate rat dorsal root ganglia neurons, although their effectiveness was lower in comparison to primary rat SCs. In toxicity assays, immortalized hSCs remain susceptible to oxidative stress induced by H 2O 2. This study shows that, using specific immortalization techniques, it is possible to establish hSC lines that retain characteristics of typical primary hSCs. These cells are particularly useful for drug screening and studies aimed at disease mechanisms involving SCs.

Original languageEnglish (US)
Pages (from-to)423-431
Number of pages9
JournalStem Cells and Development
Volume21
Issue number3
DOIs
StatePublished - Feb 10 2012

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Schwann Cells
Phenotype
Extracellular Matrix
Oxidative Stress
Cell Culture Techniques
Polyomavirus Transforming Antigens
Neurons
Preclinical Drug Evaluations
Growth Factor Receptors
Viral Tumor Antigens
Spinal Ganglia
Peripheral Nervous System Diseases
Coculture Techniques
Transcriptome
Neuroglia
Reverse Transcription
Axons
Organism Cloning
Clone Cells
Immunohistochemistry

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Hematology

Cite this

Human schwann cells retain essential phenotype characteristics after immortalization. / Lehmann, Helmar C.; Chen, Weiran; Mi, Ruifa; Wang, Shuo; Liu, Ying; Rao, Mahendra; Hoke, Ahmet.

In: Stem Cells and Development, Vol. 21, No. 3, 10.02.2012, p. 423-431.

Research output: Contribution to journalArticle

Lehmann, Helmar C. ; Chen, Weiran ; Mi, Ruifa ; Wang, Shuo ; Liu, Ying ; Rao, Mahendra ; Hoke, Ahmet. / Human schwann cells retain essential phenotype characteristics after immortalization. In: Stem Cells and Development. 2012 ; Vol. 21, No. 3. pp. 423-431.
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