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Morphological, molecular and FTIR spectroscopic analysis during the differentiation of kidney cells from pluripotent
Monica Maribel Mata-Miranda1,2, Gustavo Jesus Vazquez-Zapien1,3, Marlon Rojas-Lopez1
1Centro de Investigación en Biotecnología Aplicada, CIBA-Tlaxcala, Instituto Politécnico Nacional, 90700, Tepetitla, Tlaxcala, Mexico.
Background:
Kidney diseases are a global health problem. Currently, over 2 million people require dialysis or transplant which are associated with high morbidity and mortality; therefore, new researches focused on regenerative medicine have been developed, including the use of stem cells.
Results:
In this research, we generate differentiated kidney cells (DKCs) from mouse pluripotent stem cells (mPSCs) analyzing their morphological, genetic, phenotypic, and spectroscopic characteristics along differentiation, highlighting that there are no reports of the use of Fourier transform infrared (FTIR) spectroscopy to characterize the directed differentiation of mPSCs to DKCs. The genetic and protein experiments proved the obtention of DKCs that passed through the chronological stages of embryonic kidney development. Regarding vibrational spectroscopy analysis by FTIR, bands related with biomolecules were shown on mPSCs and DKCs spectra, observing distinct differences between cell lineages and maturation stages. The second derivative of DKCs spectra showed changes in the protein bands compared to mPSCs. Finally, the principal components analysis obtained from FTIR spectra allowed to characterize chemical and structurally mPSCs and their differentiation process to DKCs in a rapid and non-invasive way.
Conclusion:
Our results indicated that we obtained DKCs from mPSCs, which passed through the chronological stages of embryonic kidney development. Moreover, FTIR spectroscopy resulted in a non-invasive, rapid and precise technic that together with principal component analysis allows to characterize chemical and structurally both kind of cells and also discriminate and determine different stages along the cell differentiation process.
Insights
Researchers successfully generated differentiated kidney cells (DKCs) from mouse pluripotent stem cells (mPSCs). Fourier transform infrared (FTIR) spectroscopy rapidly and non-invasively characterized this stem cell differentiation process.
Area of Science:
- Regenerative Medicine
- Stem Cell Biology
- Biotechnology
Background:
- Kidney diseases pose a significant global health challenge, necessitating advanced treatments.
- Current treatments like dialysis and transplantation have high morbidity and mortality rates.
- Regenerative medicine, particularly stem cell therapy, offers promising avenues for kidney disease research.
Purpose of the Study:
- To generate differentiated kidney cells (DKCs) from mouse pluripotent stem cells (mPSCs).
- To characterize the differentiation process using morphological, genetic, phenotypic, and spectroscopic methods.
- To explore the utility of Fourier transform infrared (FTIR) spectroscopy for characterizing stem cell differentiation into DKCs.
Main Methods:
- Directed differentiation of mouse pluripotent stem cells (mPSCs) into differentiated kidney cells (DKCs).
- Analysis of cell characteristics including morphology, genetics, and phenotype.
- Application of Fourier transform infrared (FTIR) spectroscopy and principal component analysis (PCA) for spectroscopic characterization.
Main Results:
- DKCs were successfully generated from mPSCs, recapitulating embryonic kidney development stages.
- FTIR spectroscopy revealed distinct spectral differences between mPSCs and DKCs, indicating changes in biomolecular composition.
- PCA of FTIR spectra enabled rapid, non-invasive chemical and structural characterization of mPSCs and their differentiation into DKCs.
Conclusions:
- The study successfully produced DKCs from mPSCs, mirroring embryonic kidney development.
- FTIR spectroscopy, coupled with PCA, provides a rapid, non-invasive, and precise method for characterizing stem cell differentiation.
- This spectroscopic approach can discriminate between cell types and monitor differentiation stages effectively.