Defining optimal enzyme and matrix combination for replating of human induced pluripotent stem cell-derived cardiomyocytes at different levels of maturity

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dc.authorid0000-0002-4022-1014
dc.authorid0000-0002-7199-583X
dc.contributor.authorKoç, Arzuhan
dc.contributor.authorŞahoğlu Göktaş, Sevilay
dc.contributor.authorAkgül Çağlar, Tuba
dc.contributor.authorÇağavi, Esra
dc.date.accessioned2021-05-21T08:09:52Z
dc.date.available2021-05-21T08:09:52Z
dc.date.issued2021
dc.departmentİstanbul Medipol Üniversitesi, Rektörlük, Rejeneratif ve Restoratif Tıp Araştırmaları Merkezi (REMER)
dc.departmentİstanbul Medipol Üniversitesi, Rektörlük, Sağlık Bilim ve Teknolojileri Araştırma Enstitüsü
dc.departmentİstanbul Medipol Üniversitesi, Sağlık Bilimleri Enstitüsü, Mikrobiyoloji Ana Bilim Dalı
dc.departmentİstanbul Medipol Üniversitesi, Sağlık Bilimleri Enstitüsü, Sinirbilim Ana Bilim Dalı
dc.departmentİstanbul Medipol Üniversitesi, Sağlık Bilimleri Enstitüsü, Tıbbi Biyoloji ve Genetik Ana Bilim Dalı
dc.departmentİstanbul Medipol Üniversitesi, Tıp Fakültesi, Temel Tıp Bilimleri Bölümü, Tıbbi Biyoloji Ana Bilim Dalı
dc.description.abstractHuman induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) create an unlimited cell source for basic and translational research. Depending on the maturity of cardiac cultures and the intended applications, obtaining hiPSC-CMs as a single-cell, monolayer or three-dimensional clusters can be challenging. Here, we defined strategies to replate hiPSC-CMs on early days (D15-30) or later more mature (D60-150) differentiation cultures. After generation of hiPSCs and derivation of cardiomyocytes, four dissociation reagents Collagenase A/B, Collagenase II, TrypLE, EDTA and five different extracellular matrix materials Laminin, iMatrix-511, Fibronectin, Matrigel, and Geltrex were comparatively evaluated by imaging, cell viability, and contraction analysis. For early cardiac differentiation cultures mimicking mostly the embryonic stage, the highest adhesion, cell viability, and beating frequencies were achieved by treatment with the TrypLE enzyme. Video-based contraction analysis demonstrated higher beating rates after replating compared to before treatment. For later differentiation days of more mature cardiac cultures, dissociation with EDTA and replating cells on Geltrex or Laminin-derivatives yielded better recovery. Cardiac clusters at various sizes were detected in several groups treated with collagenases. Collectively, our findings revealed the selection criteria of the dissociation approach and coating matrix for replating iPSC-CMs based on the maturity and the requirements of further downstream applications.
dc.identifier.citationKoç, A., Şahoğlu Göktaş, S., Akgül Çağlar, T. ve Çağavi, E. (2021). Defining optimal enzyme and matrix combination for replating of human induced pluripotent stem cell-derived cardiomyocytes at different levels of maturity. Experimental Cell Research, 403(2). https://dx.doi.org/10.1016/j.yexcr.2021.112599
dc.identifier.doi10.1016/j.yexcr.2021.112599
dc.identifier.issn0014-4827
dc.identifier.issn1090-2422
dc.identifier.issue2
dc.identifier.scopusqualityQ2
dc.identifier.urihttps://dx.doi.org/10.1016/j.yexcr.2021.112599
dc.identifier.urihttps://hdl.handle.net/20.500.12511/6874
dc.identifier.volume403
dc.identifier.wosqualityQ3
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherElsevier Inc.
dc.relation.ispartofExperimental Cell Researchen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.relation.tubitakinfo:eu-repo/grantAgreement/TUBITAK/SOBAG/119S132
dc.relation.tubitakinfo:eu-repo/grantAgreement/TUBITAK/SOBAG/213S192
dc.rightsinfo:eu-repo/semantics/embargoedAccess
dc.subjectHuman Induced Pluripotent Stem Cells
dc.subjectCardiomyocytes
dc.subjectCardiac Differentiation
dc.subjectReplating
dc.subjectCell Dissociation
dc.subjectDissociation Enzymes
dc.subjectExtracellular Matrix
dc.titleDefining optimal enzyme and matrix combination for replating of human induced pluripotent stem cell-derived cardiomyocytes at different levels of maturity
dc.typeArticle

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