RRC ID 82051
Author Urasawa T, Koizumi T, Kimura K, Ohta Y, Kawasaki N.
Title Quantitative Proteomics for the Development and Manufacturing of Human-Induced Pluripotent Stem Cell-Derived Neural Stem Cells Using Data-Independent Acquisition Mass Spectrometry.
Journal J Proteome Res
Abstract Human-induced pluripotent stem cell (iPSC)-derived neural stem cells (NSCs) have several potential applications in regenerative medicine. A deep understanding of stem cell characteristics is critical for developing appropriate products for use in the clinic. This study aimed to develop approaches for characterizing iPSC-derived NSCs. Data-independent acquisition mass spectrometry (DIA-MS) was used to obtain temporal proteomic profiles of differentiating cells. Principal component analysis of the proteome profiles allowed for the discrimination of cells cultured for different periods. Cells were characterized by Gene Ontology analysis to annotate the upregulated proteins based on their functions. We found that trophoblast glycoprotein (TPBG), a membrane glycoprotein that inhibits the Wnt/β-catenin pathway, was elevated in NSC and that silencing TPBG promoted proliferation rather than neuronal differentiation. Treatment with Wnt/β-catenin pathway activators and inhibitors showed that modulating the Wnt/β-catenin pathway is crucial for differentiation into NSC. These results suggest that the level of TPBG is critical for differentiation into NSC, and TPBG is a potentially critical quality attribute of differentiating cells. In summary, DIA-MS-based proteomics is a promising multi-attribute method for characterizing stem cell-derived products.
Volume 22(6)
Pages 1843-1854
Published 2023-6-2
DOI 10.1021/acs.jproteome.2c00841
PMID 37097202
PMC PMC10243303
MeSH Cell Differentiation Humans Induced Pluripotent Stem Cells* / metabolism Neural Stem Cells* Proteomics Wnt Signaling Pathway beta Catenin / genetics beta Catenin / metabolism
Resource
Human and Animal Cells 201B7(HPS0063) 610B1(HPS0331)