Simultaneous and quantitative monitoring transcription factors in human embryonic stem cell differentiation using mass spectrometry–based targeted proteomics

Xu, Mengying; Xu, Lei; Cao, Jianxiang; Hu, Yechen; Xu, Feifei; Liu, Yan; Chen, Yun

doi:10.1007/s00216-021-03160-7

Cited by 1 publication

(1 citation statement)

References 43 publications

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“…3 , the iPSCs exhibited strong nuclear expression of Oct4 and SOX2, with almost no expression of Nestin, which is localized to the cytoskeleton. This aligns with the characteristic features of iPSCs, where SOX2 and Oct4, key regulators maintaining pluripotency, are localized within the nucleus [ 34 ]. In contrast, in NPCs, SOX2 and Oct4 were not expressed at all, whereas the cytoskeletal proteins Nestin and the nuclear-localised PAX6 were strongly expressed.…”

Section: Resultssupporting

confidence: 76%

DHM/SERS reveals cellular morphology and molecular changes during iPSCs-derived activation of astrocytes

Bu,

Yang,

Han

et al. 2024

Biomed. Opt. Express

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The activation of astrocytes derived from induced pluripotent stem cells (iPSCs) is of great significance in neuroscience research, and it is crucial to obtain both cellular morphology and biomolecular information non-destructively in situ, which is still complicated by the traditional optical microscopy and biochemical methods such as immunofluorescence and western blot. In this study, we combined digital holographic microscopy (DHM) and surface-enhanced Raman scattering (SERS) to investigate the activation characteristics of iPSCs-derived astrocytes. It was found that the projected area of activated astrocytes decreased by 67%, while the cell dry mass increased by 23%, and the cells changed from a flat polygonal shape to an elongated star-shaped morphology. SERS analysis further revealed an increase in the intensities of protein spectral peaks (phenylalanine 1001 cm-1, proline 1043 cm-1, etc.) and lipid-related peaks (phosphatidylserine 524 cm-1, triglycerides 1264 cm-1, etc.) decreased in intensity. Principal component analysis-linear discriminant analysis (PCA-LDA) modeling based on spectral data distinguished resting and reactive astrocytes with a high accuracy of 96.5%. The increase in dry mass correlated with the increase in protein content, while the decrease in projected area indicated the adjustment of lipid composition and cell membrane remodeling. Importantly, the results not only reveal the cellular morphology and molecular changes during iPSCs-derived astrocytes activation but also reflect their mapping relationship, thereby providing new insights into diagnosing and treating neurodegenerative diseases.

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Section: Resultssupporting

confidence: 76%