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Both infrared (IR) and Raman microspectroscopies have been applied, with a clear predominance of the latter. Bone marrow MSC have been the target of most studies, followed by those originating from dental\/oral and adipose tissues. Interests have increasingly addressed single cell and extracellular matrix characterization at the molecular level. Most studies have focused on the characteristics and maturity of time-course mineralization, attempting to localize mineral aggregates formed onto the evolving collagen strands. Some reports have focused on time-dependent changes in protein structure and other components of extracellular matrix components. Besides spectral band examination through position, linewidth and shape, selected band ratios have proved largely informative to assess mineral species evolution and mineral-to-organic matrix interactions over time. The increasing use of multivariate analysis (or chemometrics) and machine learning strategies to detect finer spectral variations is evident, as is the promise of more recent IR and Raman variations to provide higher sensitivity and spatial resolution conditions. The label-free non-invasive nature of vibrational microspectroscopy makes it particularly promising for rapid and effective selection of suitable MSC donors, to support scale-up procedures for translation to the clinic. Some of the challenges to be faced are briefly discussed.<\/jats:p>\n          <jats:p>\n            <jats:bold>Graphical Abstract<\/jats:bold>\n          <\/jats:p>","DOI":"10.1007\/s12015-025-10943-3","type":"journal-article","created":{"date-parts":[[2025,7,26]],"date-time":"2025-07-26T05:46:40Z","timestamp":1753508800000},"page":"2043-2065","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Exploring In Vitro Mesenchymal Stem Cell Osteodifferentiation via Vibrational Microspectroscopy: A Review"],"prefix":"10.1007","volume":"21","author":[{"given":"Daniela S.","family":"Bispo","sequence":"first","affiliation":[]},{"given":"In\u00eas C. R.","family":"Gra\u00e7a","sequence":"additional","affiliation":[]},{"given":"Jo\u00e3o A.","family":"Rodrigues","sequence":"additional","affiliation":[]},{"given":"Jo\u00e3o T. 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