Stem Cell Reviews and Reports

, Volume 7, Issue 2, pp 394–403

Efficient Generation of Schwann Cells from Human Embryonic Stem Cell-Derived Neurospheres

  • Lina Ziegler
  • Sergei Grigoryan
  • In Hong Yang
  • Nitish V. Thakor
  • Ronald S. Goldstein
Article

Abstract

Schwann cells (SC), the glial cells of peripheral nerves, are involved in many diseases including Charcot Marie Tooth and neurofibromatosis, and play a pivotal role in peripheral nerve regeneration. Although it is possible to obtain human SC from nerve biopsies, they are difficult to maintain and expand in culture. Here we describe an efficient system for directing the differentiation of human embryonic stem cells (hESC) into cells with the morphological and molecular characteristics of SC. Neurospheres were generated from hESC using stromal cell induction and grown under conditions supportive of SC differentiation. After 8 weeks, hESC-derived SC expressed characteristic markers GFAP, S100, HNK1, P75, MBP and PMP-22, and were observed in close association with hESC-derived neurites. ~60% of the cells were double-immunostained for the SC markers GFAP/S100. RT-PCR analysis confirmed the expression of GFAP, S100, P75, PMP-22 and MBP and demonstrated expression of the SC markers P0, KROX20 and PLP in the cultures. Expression of CAD19 was observed in 2 and 4 week cultures and then was down-regulated, consistent with its expression in SC precursor, but not mature stages. Co-culture of hESC-derived SC with rat, chick or hESC-derived axons in compartmentalized microfluidic chambers resulted in tight association of the SC with axons. Apparent wrapping of the axons by SC was occasionally observed, suggestive of myelination. Our method for generating SC from hESC makes available a virtually unlimited source of human SC for studies of their role in nerve regeneration and modeling of disease.

Keywords

Human embryonic stem cells Schwann cells Myelination Microfluidic chambers Peripheral nervous system 

Supplementary material

12015_2010_9198_Fig5_ESM.gif (485 kb)
Supplementary Figure 1

Scheme depicting hESC differentiation into SC and their interaction with neurons. Neurospheres were generated using the PA6 induction method, grown in suspension for 4 wk and differentiated under adherent conditions into putative SC for a further 8 wk. In some experiments, differentiated SC were co-cultured with chick / rat or hESC-derived neurons (GIF 484 kb)

12015_2010_9198_MOESM1_ESM.tif (1.3 mb)
High resolution image (TIFF 1324 kb)
12015_2010_9198_Fig6_ESM.gif (144 kb)
Supplementary Figure 2

Oil red staining of cultures of hESC-derived SC. After fixation with 4% paraformaldehyde and rinsing with 60% isopropanol, 8-wk cultures were stained with freshly prepared Oil Red and lightly counterstained with hematoxylin. The red staining of many cells indicates high concentrations of lipids, indicative of myelination (GIF 144 kb)

12015_2010_9198_MOESM2_ESM.tif (1.8 mb)
High resolution image (TIFF 1827 kb)

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Lina Ziegler
    • 1
  • Sergei Grigoryan
    • 1
  • In Hong Yang
    • 2
  • Nitish V. Thakor
    • 2
  • Ronald S. Goldstein
    • 1
  1. 1.Mina and Everard Goodman Faculty of Life SciencesBar-Ilan UniversityRamat-GanIsrael
  2. 2.Department of Biomedical Engineering, School of MedicineJohns Hopkins UniversityBaltimoreUSA

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