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Article: Generating mESC-derived insulin-producing cell lines through an intermediate lineage-restricted progenitor line

TitleGenerating mESC-derived insulin-producing cell lines through an intermediate lineage-restricted progenitor line
Authors
Issue Date2009
PublisherELSEVIER
Citation
Stem Cell Research, 2009, v. 2 n. 1, p. 41-55 How to Cite?
AbstractGenerating surrogate insulin-producing cells from embryonic stem cells (ESCs) through in vitro replication of successive steps during pancreatic development has been challenging . Here we describe a novel reproducible protocol to establish homogeneous and scalable insulin-producing cell lines from mouse (m) ESCs via differentiation of the previously described lineage-restricted clonal mESC-derived E-RoSH cells. Unlike their parental mESCs, E-RoSH cells expressed high levels of mesodermal and endodermal genes. Nutrient depletion in the presence of nicotinamide inhibited proliferation of E-RoSH cells and induced differentiation into heterogeneous cultures comprising vascular-like structures that produced detectable levels of insulin and C-peptide in an equimolar ratio. Limiting dilution of these cultures resulted in the isolation of eight independent insulin-producing cell lines in five experiments. All these lines were cloned and shown to be amenable to repeated cycles of freeze and thaw and to replicate for months with a doubling time of 3-4 days. Under such conditions, the cultured cells exhibited genomic, structural, biochemical, and pharmacological properties of pancreatic β cells, including storage of an equimolar ratio of insulin and C-peptide in granules and release of the contents of these organelles through a glucose-sensitive machinery. After transplantation, these cells reversed hyperglycemia in streptozotocin-treated SCID mice and did not form teratomas. © 2008.
Persistent Identifierhttp://hdl.handle.net/10722/59206
ISSN
2015 Impact Factor: 3.892
2015 SCImago Journal Rankings: 2.037
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLi, Gen_HK
dc.contributor.authorLuo, Ren_HK
dc.contributor.authorZhang, Jen_HK
dc.contributor.authorYeo, KSen_HK
dc.contributor.authorLian, Qen_HK
dc.contributor.authorXie, Fen_HK
dc.contributor.authorTan, EKWen_HK
dc.contributor.authorCaille, Den_HK
dc.contributor.authorKon, OLen_HK
dc.contributor.authorSaltoTellez, Men_HK
dc.contributor.authorMeda, Pen_HK
dc.contributor.authorLim, SKen_HK
dc.date.accessioned2010-05-31T03:45:06Z-
dc.date.available2010-05-31T03:45:06Z-
dc.date.issued2009en_HK
dc.identifier.citationStem Cell Research, 2009, v. 2 n. 1, p. 41-55en_HK
dc.identifier.issn1873-5061en_HK
dc.identifier.urihttp://hdl.handle.net/10722/59206-
dc.description.abstractGenerating surrogate insulin-producing cells from embryonic stem cells (ESCs) through in vitro replication of successive steps during pancreatic development has been challenging . Here we describe a novel reproducible protocol to establish homogeneous and scalable insulin-producing cell lines from mouse (m) ESCs via differentiation of the previously described lineage-restricted clonal mESC-derived E-RoSH cells. Unlike their parental mESCs, E-RoSH cells expressed high levels of mesodermal and endodermal genes. Nutrient depletion in the presence of nicotinamide inhibited proliferation of E-RoSH cells and induced differentiation into heterogeneous cultures comprising vascular-like structures that produced detectable levels of insulin and C-peptide in an equimolar ratio. Limiting dilution of these cultures resulted in the isolation of eight independent insulin-producing cell lines in five experiments. All these lines were cloned and shown to be amenable to repeated cycles of freeze and thaw and to replicate for months with a doubling time of 3-4 days. Under such conditions, the cultured cells exhibited genomic, structural, biochemical, and pharmacological properties of pancreatic β cells, including storage of an equimolar ratio of insulin and C-peptide in granules and release of the contents of these organelles through a glucose-sensitive machinery. After transplantation, these cells reversed hyperglycemia in streptozotocin-treated SCID mice and did not form teratomas. © 2008.en_HK
dc.languageengen_HK
dc.publisherELSEVIERen_HK
dc.relation.ispartofStem Cell Researchen_HK
dc.subject.meshAnimalsen_HK
dc.subject.meshC-Peptide - analysisen_HK
dc.subject.meshCell Culture Techniquesen_HK
dc.subject.meshCell Differentiationen_HK
dc.subject.meshCell Lineageen_HK
dc.subject.meshCell Transplantationen_HK
dc.subject.meshEmbryonic Stem Cells - cytologyen_HK
dc.subject.meshEndodermen_HK
dc.subject.meshHyperglycemia - therapyen_HK
dc.subject.meshInsulin - analysisen_HK
dc.subject.meshInsulin-Secreting Cells - cytologyen_HK
dc.subject.meshMesodermen_HK
dc.subject.meshMiceen_HK
dc.subject.meshMice, SCIDen_HK
dc.subject.meshTreatment Outcomeen_HK
dc.titleGenerating mESC-derived insulin-producing cell lines through an intermediate lineage-restricted progenitor lineen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1873-5061&volume=2009 Jan;2&issue=1&spage=41&epage=55&date=2009&atitle=Generating+mESC-derived+insulin-producing+cell+lines+through+an+intermediate+lineage-restricted+progenitor+line.en_HK
dc.identifier.emailLian, Q:qzlian@hkucc.hku.hken_HK
dc.identifier.authorityLian, Q=rp00267en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.scr.2008.07.006en_HK
dc.identifier.pmid19383408en_HK
dc.identifier.scopuseid_2-s2.0-56249130373en_HK
dc.identifier.hkuros161065en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-56249130373&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume2en_HK
dc.identifier.issue1en_HK
dc.identifier.spage41en_HK
dc.identifier.epage55en_HK
dc.identifier.isiWOS:000269909000006-
dc.identifier.scopusauthoridLi, G=10739742900en_HK
dc.identifier.scopusauthoridLuo, R=7202671143en_HK
dc.identifier.scopusauthoridZhang, J=12781492000en_HK
dc.identifier.scopusauthoridYeo, KS=22952467200en_HK
dc.identifier.scopusauthoridLian, Q=7003399023en_HK
dc.identifier.scopusauthoridXie, F=35309079600en_HK
dc.identifier.scopusauthoridTan, EKW=14012680600en_HK
dc.identifier.scopusauthoridCaille, D=6701613373en_HK
dc.identifier.scopusauthoridKon, OL=7004607001en_HK
dc.identifier.scopusauthoridSaltoTellez, M=7003434917en_HK
dc.identifier.scopusauthoridMeda, P=7005822187en_HK
dc.identifier.scopusauthoridLim, SK=7404080956en_HK

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