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Raman microspectroscopy fingerprinting of organoid differentiation state
Kate Tubbesing1,2,3,4, Nicholas Moskwa2,3,5, Ting Chean Khoo1
1Department of Physics, University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY, 12222, USA.
Background:
Organoids, which are organs grown in a dish from stem or progenitor cells, model the structure and function of organs and can be used to define molecular events during organ formation, model human disease, assess drug responses, and perform grafting in vivo for regenerative medicine approaches. For therapeutic applications, there is a need for nondestructive methods to identify the differentiation state of unlabeled organoids in response to treatment with growth factors or pharmacologicals.
Methods:
Using complex 3D submandibular salivary gland organoids developed from embryonic progenitor cells, which respond to EGF by proliferating and FGF2 by undergoing branching morphogenesis and proacinar differentiation, we developed Raman confocal microspectroscopy methods to define Raman signatures for each of these organoid states using both fixed and live organoids.
Results:
Three separate quantitative comparisons, Raman spectral features, multivariate analysis, and machine learning, classified distinct organoid differentiation signatures and revealed that the Raman spectral signatures were predictive of organoid phenotype.
Conclusions:
As the organoids were unlabeled, intact, and hydrated at the time of imaging, Raman spectral fingerprints can be used to noninvasively distinguish between different organoid phenotypes for future applications in disease modeling, drug screening, and regenerative medicine.
