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{"title":"Label-FreenRaman Spectroscopy for Assessing Puritynand Maturation of hiPSC-Derived Cardiac Tissue","authors":"Junjun Li (282515), Menglu Li (4458208), Yasunori Nawa (3953441), Yuting Liu (270221), Kazuki Bando (4656982), Ying Hua (452923), Lifu Sun (118982), Satoshi Fujita (545706), Yoshiki Sawa (219191), Katsumasa Fujita (506421), Li Liu (75607)","doi":"10.1021/acs.analchem.4c03871.s001","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c03871.s001","url":null,"abstract":"I. Background: Humanninduced pluripotent stem cell (hiPSC) derivedncardiomyocytes (CMs) have been utilized in drug toxicity evaluation,ndrug discovery, and treating heart failure patients, showing substantialneffects. Ensuring the quality, purity, and maturation of hiPSC-CMsnduring large-scale production is crucial. There is a growing demandnfor a novel method to characterize cell molecular profiles withoutnlabels and without causing damage. II. Methods: In this study, wenemployed label-free Raman microscopy to evaluate hiPSC-derived CMs.nThe study involved the characterization of cell molecular profilesnwithout labels and without causing damage. The correlation betweennRaman spectroscopy of specific components, such as cytochrome <i>c</i> and myoglobin, and CM purity and maturation followingnhiPSC differentiation was investigated. Additionally, the validationnof this correlation was performed by assessing mixtures of commerciallynavailable CMs (iCell cardiomyocytes2) and fibroblasts at various ratiosnas well as hiPSC-derived CMs with different efficiencies. Furthermore,nCMs were matured using rapid pacing of traveling waves, and the Ramannprofiles of matured CMs were compared to those of immature ones. III.nResults: Raman spectroscopy indicated that the cytochrome <i>c</i> and myoglobin showed correlation with the purity and maturationnof CMs following differentiation of hiPSCs. This correlation was validatednthrough experiments involving different CM-fibroblast mixtures andnhiPSC-derived CMs with varying efficiencies. Moreover, matured CMsnexhibited markedly different Raman profiles compared to immature ones,nindicating the potential of Raman imaging as a tool for assessingnCM maturation. IV. Conclusions: We discovered that Raman spectroscopynof certain components, such as cytochrome <i>c</i> and myoglobin,ncorrelates with the CM purity and maturation following hiPSC differentiation.nThe findings of this study highlight the potential of label-free Ramannmicroscopy as a nondestructive, high-content, and time-efficient methodnfor quality control of hiPSC-derived CMs. This approach could significantlyncontribute to ensuring the quality and maturity of hiPSC-CMs for variousnapplications in drug discovery and regenerative medicine.","PeriodicalId":521900,"journal":{"name":"Figshare","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147887552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}