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Conference Paper: Study of extracellular matrix for the culture of human embryonic stem cells

TitleStudy of extracellular matrix for the culture of human embryonic stem cells
Authors
Issue Date2009
PublisherInternational Society For Stem Cell Research.
Citation
The 7th Annual Meeting of the International Society For Stem Cell Research (ISSCR 2009), Barcelona Spain, 8-11 July 2009, p. 113 How to Cite?
AbstractBackground: The development of clinical grade human embryonic stem cells (hESC) suitable for cell therapy requires defined components for hESC culture. We hypothesize that the feeder cells used for hESC culture produce extracellular matrix molecules supporting the growth of hESC. Materials and Methods: The profiles of mRNA and extracellular matrix production of two human feeder cell lines, human foreskin fibroblast (hFF-1) and human lung fibroblast (WI-38) were correlated with their ability to maintain the pluripotency of a hESC line, BG01V. The mRNA expression profile and matrix production were determined by Affymetrix microarray chip and liquid chromatography/mass spectrometry/mass spectrometry analysis, respectively. Results: Using a morphological grading system, hFF-1 but not WI-38 supported the pluripotency of BG01V. The observation was supported by over-expression of early differentiation markers KRT-8 and -18 mRNA in BG01V cells cultured on WI-38, though the mRNA expression of pluripotent markers Nanog, Oct4 and TRA-1-81 in BG01V cultured with hFF-1 and WI-38 was similar. Microarray analysis showed that the expression of 530 transcripts was 2-fold higher (P<0.05) in hFF-1 than in WI-38. The microarray data were validated by real-time PCR on 12 differentially expressed genes related to secretory and extracellular matrix molecules. Proteomic profiling of the extracellular matrix molecules also demonstrated differences between the two feeder cell lines. Both microarray and proteomic analysis identified sulfatase 1 (SUFL1) and chemokine (C-X-C Motif) ligand 12 (CXCL12) were higher in hFF-1. Conclusion: The difference in the ability of feeder cells in maintaining the pleuripotency of hESC may be related to the differential secretion of matrix molecules by the feeder cells.
DescriptionPoster Session 2: abstract no. 718
Persistent Identifierhttp://hdl.handle.net/10722/113632

 

DC FieldValueLanguage
dc.contributor.authorLee, CYLen_HK
dc.contributor.authorHou, YKen_HK
dc.contributor.authorPang, RTKen_HK
dc.contributor.authorLee, CKFen_HK
dc.contributor.authorYeung, WSBen_HK
dc.date.accessioned2010-09-26T04:24:23Z-
dc.date.available2010-09-26T04:24:23Z-
dc.date.issued2009en_HK
dc.identifier.citationThe 7th Annual Meeting of the International Society For Stem Cell Research (ISSCR 2009), Barcelona Spain, 8-11 July 2009, p. 113-
dc.identifier.urihttp://hdl.handle.net/10722/113632-
dc.descriptionPoster Session 2: abstract no. 718-
dc.description.abstractBackground: The development of clinical grade human embryonic stem cells (hESC) suitable for cell therapy requires defined components for hESC culture. We hypothesize that the feeder cells used for hESC culture produce extracellular matrix molecules supporting the growth of hESC. Materials and Methods: The profiles of mRNA and extracellular matrix production of two human feeder cell lines, human foreskin fibroblast (hFF-1) and human lung fibroblast (WI-38) were correlated with their ability to maintain the pluripotency of a hESC line, BG01V. The mRNA expression profile and matrix production were determined by Affymetrix microarray chip and liquid chromatography/mass spectrometry/mass spectrometry analysis, respectively. Results: Using a morphological grading system, hFF-1 but not WI-38 supported the pluripotency of BG01V. The observation was supported by over-expression of early differentiation markers KRT-8 and -18 mRNA in BG01V cells cultured on WI-38, though the mRNA expression of pluripotent markers Nanog, Oct4 and TRA-1-81 in BG01V cultured with hFF-1 and WI-38 was similar. Microarray analysis showed that the expression of 530 transcripts was 2-fold higher (P<0.05) in hFF-1 than in WI-38. The microarray data were validated by real-time PCR on 12 differentially expressed genes related to secretory and extracellular matrix molecules. Proteomic profiling of the extracellular matrix molecules also demonstrated differences between the two feeder cell lines. Both microarray and proteomic analysis identified sulfatase 1 (SUFL1) and chemokine (C-X-C Motif) ligand 12 (CXCL12) were higher in hFF-1. Conclusion: The difference in the ability of feeder cells in maintaining the pleuripotency of hESC may be related to the differential secretion of matrix molecules by the feeder cells.-
dc.languageengen_HK
dc.publisherInternational Society For Stem Cell Research.-
dc.relation.ispartofAnnual Meeting of the International Society For Stem Cell Research, ISSCR 2009-
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleStudy of extracellular matrix for the culture of human embryonic stem cellsen_HK
dc.typeConference_Paperen_HK
dc.identifier.emailLee, CYL: cherielee@hku.hken_HK
dc.identifier.emailPang, RTK: rtkpang@hku.hken_HK
dc.identifier.emailLee, CKF: ckflee@hku.hken_HK
dc.identifier.emailYeung, WSB: wsbyeung@hku.hken_HK
dc.identifier.authorityLee, CYL=rp00308en_HK
dc.identifier.authorityPang, RTK=rp01761en_HK
dc.identifier.authorityLee, CKF=rp00458en_HK
dc.description.naturepublished_or_final_version-
dc.identifier.hkuros163982en_HK
dc.identifier.spage113-
dc.identifier.epage113-
dc.publisher.placeUnited States-

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