Evaluation of Differentiated/Undifferentiated State of iPS Cells Using DPiMS™-2020 and eMSTAT Solution™

Importance of the Topic

Pluripotent stem cells are central to regenerative medicine research and quality control. Rapid, minimally invasive evaluation of their differentiated or undifferentiated status is critical to optimize cell culture protocols, conserve valuable samples, and accelerate decision making in both academic and industrial settings.

Objectives and Study Overview

This application note demonstrates a streamlined workflow for assessing the differentiation state of human induced pluripotent stem (iPS) cells by analyzing the composition of spent culture media. The study combined probe electro spray ionization on the DPiMS-2020 mass spectrometer with multivariate statistical analysis in eMSTAT Solution to distinguish media from undifferentiated, spheroid-differentiated, and adherent-differentiated cells.

Methodology

iPS cells were grown under defined conditions for 2 and 7 days in Essential8™ with or without fetal bovine serum (FBS) and gelatin coating. Spent media supernatants were mixed 1:1 with ethanol. Ten microliters of each sample were deposited onto a specialized liquid sample plate and directly analyzed in scan mode, generating mass spectra within approximately three minutes per sample.

Instrumentation Used

DPiMS-2020 mass spectrometer employing probe electro spray ionization for direct media analysis
eMSTAT Solution software for multivariate statistical evaluation

Main Results and Discussion

Distinct spectral profiles allowed clear separation of undifferentiated and differentiated culture media. Score plots revealed that samples cluster according to both differentiation state and progression (day 2 vs. day 7). Overlap between adherent day-2 and spheroid day-7 groups indicated similar differentiation levels, highlighting the method’s sensitivity to subtle metabolic changes.

Benefits and Practical Applications

The combined approach achieves rapid, label-free assessment with minimal sample consumption and no cell fractionation. It supports high-throughput monitoring of culture conditions, streamlines cell quality control workflows, and reduces reliance on more laborious molecular assays.

Future Trends and Possibilities

Future developments may include integration with automated sampling systems for real-time monitoring, expansion to other cell types and culture formats, and incorporation of machine learning algorithms to enhance predictive accuracy.

Conclusion

Probe electro spray ionization on the DPiMS-2020, coupled with eMSTAT multivariate analysis, provides a fast, simple, and effective strategy for evaluating iPS cell differentiation via culture media profiling. This method promises to improve throughput and reduce sample consumption in regenerative medicine research and quality control workflows.