Skip to Content
Merck
  • Mutant astrocytes differentiated from Rett syndrome patients-specific iPSCs have adverse effects on wild-type neurons.

Mutant astrocytes differentiated from Rett syndrome patients-specific iPSCs have adverse effects on wild-type neurons.

Human molecular genetics (2014-01-15)
Emily Cunningham Williams, Xiaofen Zhong, Ahmed Mohamed, Ronghui Li, Yan Liu, Qiping Dong, Gene E Ananiev, Jonathan Chern Choong Mok, Benjamin Ray Lin, Jianfeng Lu, Cassandra Chiao, Rachel Cherney, Hongda Li, Su-Chun Zhang, Qiang Chang
ABSTRACT

The disease mechanism of Rett syndrome (RTT) is not well understood. Studies in RTT mouse models have suggested a non-cell-autonomous role for astrocytes in RTT pathogenesis. However, it is not clear whether this is also true for human RTT astrocytes. To establish an in vitro human RTT model, we previously generated isogenic induced pluripotent stem cell (iPSC) lines from several RTT patients carrying different disease-causing mutations. Here, we show that these RTT iPSC lines can be efficiently differentiated into astroglial progenitors and glial fibrillary acidic protein-expressing (GFAP(+)) astrocytes that maintain isogenic status, that mutant RTT astrocytes carrying three different RTT mutations and their conditioned media have adverse effects on the morphology and function of wild-type neurons and that the glial effect on neuronal morphology is independent of the intrinsic neuronal deficit in mutant neurons. Moreover, we show that both insulin-like growth factor 1 (IGF-1) and GPE (a peptide containing the first 3 amino acids of IGF-1) are able to partially rescue the neuronal deficits caused by mutant RTT astrocytes. Our findings confirm the critical glial contribution to RTT pathology, reveal potential cellular targets of IGF-1 therapy and further validate patient-specific iPSCs and their derivatives as valuable tools to study RTT disease mechanism.

MATERIALS
Product Number
Brand
Product Description

Supelco
2-Mercaptoethanol, for HPLC derivatization, LiChropur, ≥99.0% (GC)
Sigma-Aldrich
Anti-Glial Fibrillary Acidic Protein Antibody, clone GA5, clone GA5, Chemicon®, from mouse
Sigma-Aldrich
Anti-TFCP2 antibody produced in mouse, purified immunoglobulin, buffered aqueous solution
Sigma-Aldrich
Anti-GABA antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
2-Mercaptoethanol, ≥99.0%
Sigma-Aldrich
2-Mercaptoethanol, for molecular biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)
Sigma-Aldrich
2-Mercaptoethanol, BioUltra, for molecular biology, ≥99.0% (GC)
Sigma-Aldrich
FGF-2 human, recombinant, expressed in insect cells, ≥85% (SDS-PAGE)
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Heparin sodium salt from porcine intestinal mucosa, Grade I-A, ≥180 USP units/mg, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
FGF-2 human, recombinant, expressed in E. coli, ≥95% (SDS-PAGE), ≥95% (HPLC)