Spectroscopic signature of mouse embryonic stem cell–derived hepatocytes using synchrotron Fourier transform infrared microspectroscopy

Abstract

Stem cell-based therapy for liver regeneration has been proposed to overcome the persistent shortage in the supply of suitable donor organs. A requirement for this to succeed is to find a rapid method to detect functional hepatocytes, differentiated from embryonic stem cells. We propose Fourier transform infrared (FTIR) microspectroscopy as a versatile method to identify the early and last stages of the differentiation process leading to the formation of hepatocytes. Using synchrotron-FTIR microspectroscopy, the means of identifying hepatocytes at the single-cell level is possible and explored. Principal component analysis and subsequent partial least-squares (PLS) discriminant analysis is applied to distinguish endoderm induction from hepatic progenitor cells and matured hepatocyte-like cells. The data are well modeled by PLS with endoderm induction, hepatic progenitor cells, and mature hepatocyte-like cells able to be discriminated with very high sensitivity and specificity. This method provides a practical tool to monitor endoderm induction and has the potential to be applied for quality control of cell differentiation leading to hepatocyte formation.

The Characterisation of Pluripotent and Multipotent Stem Cells Using Fourier Transform Infrared Microspectroscopy

“…In another successful application of FTIR microspectroscopy to discriminate different types of differentiated mESCs, synchrotron FTIR microspectroscopy was implemented to study the differentiation of stem cells into hepatocyte-like cells [34]. The differentiation of mouse embryonic stem cells into hepatocyte-like cells is a complex, multistage process, which first consists of endoderm induction, followed by hepatic initiation, and finally, maturation into hepatocyte-like cells.…”

“…The observed increase in lipid content during hepatocyte differentiation differed from what was found in previously described work by Thumanu et al . [34] who studied the hepatogenesis of mouse embryonic stem cells and observed higher lipid levels in the progenitor cells. These disparities could have been attributed to differences in lipid signaling between the two stem cells types.…”

“…For instance, in the hepatocyte differentiation studies performed by Thumanu et al . [34], and human embryonic stem cell differentiation work carried out by Heraud et al . [32], lipid depletion was found to co-occur with a loss of pluripotency.…”

Multivariate Analysis for Fourier Transform Infrared Spectra of Complex Biological Systems and Processes

“…Alternatively, the PLS method can be used instead of PCA [6,73,74]. For instance, Sandt and colleagues, using synchrotron infrared microspectroscopy coupled with PLS-DA, were able to characterize the metabolic fingerprint of induced pluripotent stem cells (iPSCs).…”

Comparative evaluation of differential laser-induced perturbation spectroscopy as a technique to discriminate emerging skin pathology

“…32 The component loadings from the first principal component were used to assess which spectral regions best explain variations within and between data sets. 33 Component loadings were squared to facilitate direct comparison of loading across wavelength regions between sub-populations of the data set (this casts each wavelength's loading value in terms of percent variance explained in the particular component). The regions of the spectrum at higher component loading levels were considered to have the highest impact on the observed differences between DMBA treated and control mice.…”