Characterization of Rhesus Macaque Embryonic Stem Cells in Primed and Naïve-like Cell States of Pluripotency Using Fourier Transform Infrared (FTIR) Microspectroscopy

TL;DR

This study uses FTIR microspectroscopy to distinguish between two states of rhesus macaque embryonic stem cells based on their biochemical profiles.

Contribution

The novel contribution is the use of FTIR microspectroscopy to non-invasively detect and classify pluripotent stem cell states in non-human primates.

Findings

FTIR spectra revealed distinct biochemical signatures between primed and naïve-like stem cell states.

PCA and PLS-DA analyses showed 100% specificity and sensitivity in classifying the cell states.

ALGöX-cultured cells showed higher nucleic acid and amide absorbance compared to FGF2/KOSR-cultured cells.

Abstract

We evaluated the potential of Fourier-transform infrared (FTIR) microspectroscopy for non-invasive biochemical profiling of rhesus macaque embryonic stem cells (rhESCs) cultured in either conventional FGF2/KOSR medium or a novel formulation, ALGöX. Cells from both conditions were analyzed by immunocytochemistry, RNA sequencing, and high-resolution FTIR profiling. Molecular marker expression patterns and transcriptional profiles revealed that rhESCs maintained in FGF2/KOSR were in the primed pluripotent state, whereas those cultured in ALGöX adopted a naïve-like state. FTIR spectra showed consistent differences in protein, lipid, and nucleic acid signatures, with ALGöX-cultured cells displaying higher amide I/II and nucleic acid absorbance and FGF2/KOSR-cultured cells exhibiting stronger lipid-associated bands. Principal component analysis (PCA) separated the two groups along PC−1 (64% variance), and partial least squares discriminant analysis (PLS-DA) classified samples with 100% specificity and 100% sensitivity. These findings demonstrate that FTIR microspectroscopy can reliably discriminate pluripotent state–specific biochemical features in non-human primate PSCs, providing a rapid and label-free approach for monitoring stem cell identity and quality.

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