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dc.contributor.authorUrbischek, Manuela
dc.contributor.authorRannikmae, Helena
dc.contributor.authorFoets, Thomas
dc.contributor.authorRavn, Katharina
dc.contributor.authorHyvönen, Marko
dc.contributor.authorde la Roche, Marc
dc.date.accessioned2019-04-15T23:30:47Z
dc.date.available2019-04-15T23:30:47Z
dc.date.issued2019-04-17
dc.identifier.issn2045-2322
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/291642
dc.description.abstractThe media formulations necessary for deriving and sustaining organoids from epithelial tissues such as prostate, colon, gastric, liver, pancreas, and others have been established. Critical components of organoid media are a set of growth factors that include R-spondins and BMP signalling antagonists such as Noggin or Gremlin 1. Currently, the practical limitations for formulating organoid media of reproducible potency and larger-scale media production that have hampered further technological applications of organoid technology include: the cost of growth factors such as R-spondins and Gremlin 1/Noggin and their production as defined specific activities free of contaminants that may affect organoid growth. Here we report the production of highly pure recombinant Gremlin 1 and R-spondin 1 from bacterial expression for use in organoid media. We detail the workflow for Gremlin 1 and R-spondin 1 expression, purification, quantification of cellular activity, quality control and use in media formulated for culturing organoids derived from a number of tissues. The development of precisely formulated, cost-effective media of defined specific activity will engender the development of novel applications for organoid technology.
dc.format.mediumElectronic
dc.languageeng
dc.publisherSpringer Science and Business Media LLC
dc.rightsAll rights reserved
dc.subjectOrganoids
dc.subjectAnimals
dc.subjectHumans
dc.subjectMice
dc.subjectBacteria
dc.subjectIntercellular Signaling Peptides and Proteins
dc.subjectCarrier Proteins
dc.subjectThrombospondins
dc.subjectBone Morphogenetic Proteins
dc.subjectRecombinant Proteins
dc.subjectCulture Media
dc.subjectCell Culture Techniques
dc.subjectGene Transfer Techniques
dc.titleOrganoid culture media formulated with growth factors of defined cellular activity.
dc.typeArticle
prism.issueIdentifier1
prism.publicationDate2019
prism.publicationNameSci Rep
prism.startingPage6193
prism.volume9
dc.identifier.doi10.17863/CAM.38800
dcterms.dateAccepted2019-04-03
rioxxterms.versionofrecord10.1038/s41598-019-42604-0
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2019-04-17
dc.contributor.orcidde la Roche, Marc [0000-0002-3757-082X]
dc.identifier.eissn2045-2322
rioxxterms.typeJournal Article/Review
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/P023681/1)
cam.issuedOnline2019-04-17
rioxxterms.freetoread.startdate2020-04-05


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