(mentions CMT) Efficient Generation of Schwann Cells from Human Embryonic Stem C

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Stem Cell Rev. 2010 Oct 30

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

Neurospheres

Ziegler L, Grigoryan S, Yang IH, Thakor NV, Goldstein RS.

Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Gonda

Building, Old Campus, 52900, Ramat-Gan, Israel.

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.