Raman microspectroscopy fingerprinting of organoid differentiation state.
Raman spectroscopy
Regenerative medicine
Salivary gland organoids
Tissue-engineered organoids
Journal
Cellular & molecular biology letters
ISSN: 1689-1392
Titre abrégé: Cell Mol Biol Lett
Pays: England
ID NLM: 9607427
Informations de publication
Date de publication:
28 Jun 2022
28 Jun 2022
Historique:
received:
06
02
2022
accepted:
24
05
2022
entrez:
28
6
2022
pubmed:
29
6
2022
medline:
1
7
2022
Statut:
epublish
Résumé
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. 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. 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. 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.
Sections du résumé
BACKGROUND
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
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
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
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.
Identifiants
pubmed: 35764935
doi: 10.1186/s11658-022-00347-3
pii: 10.1186/s11658-022-00347-3
pmc: PMC9238268
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
53Subventions
Organisme : NIDA NIH HHS
ID : R01 DA047410
Pays : United States
Organisme : NIDCR NIH HHS
ID : R01 DE027953
Pays : United States
Organisme : NIH HHS
ID : R01DE027953
Pays : United States
Organisme : NIH HHS
ID : R01DA047410
Pays : United States
Informations de copyright
© 2022. The Author(s).
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