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Extended and stable gene expression via nucleofection of MIDGE construct into adult human marrow mesenchymal stromal cells

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dc.Chemicals/CASerythropoietin, 11096-26-7; lipofectamine, 158571-62-1
dc.FundingDetailsMalaysia Toray Science Foundation Majlis Kanser Nasional
dc.FundingDetailsAcknowledgments We acknowledge the generous support from Malaysia Toray Science Foundation (MTSF), Scientex Foundation and Majlis Kanser Nasional (MAKNA). We thank the staff and fellow researchers from the Cell Banking Unit, Genetic Cancer Laboratory, Hemostasis Laboratory, Tissue Engineering Laboratory, Hematology Laboratory and Histopathology Laboratory of Universiti Kebangsaan Malaysia Medical Center for their technical guidance and assistance. We thank Professor Choo Kong Bung of Universiti Tunku Abdul Rahman for comments on the manuscript.
dc.citedby12
dc.contributor.affiliationsFaculty of Medicine and Health Sciences
dc.contributor.affiliationsUniversiti Kebangsaan Malaysia (UKM)
dc.contributor.affiliationsUniversiti Tunku Abdul Rahman (UTAR)
dc.contributor.affiliationsUniversiti Sains Islam Malaysia (USIM)
dc.contributor.authorMok P.L.en_US
dc.contributor.authorCheong S.K.en_US
dc.contributor.authorLeong C.F.en_US
dc.contributor.authorChua K.H.en_US
dc.contributor.authorAinoon O.en_US
dc.date.accessioned2024-05-29T01:59:45Z
dc.date.available2024-05-29T01:59:45Z
dc.date.issued2012
dc.description.abstractHuman mesenchymal stromal cell (hMSC) is a potential target for cell and gene therapy-based approaches against a variety of different diseases. Whilst cationic lipofection has been widely experimented, the Nucleofector technology is a relatively new non-viral transfection method designed for primary cells and hard-to-transfect cell lines. Herein, we compared the efficiency and viability of nucleofection with cationic lipofection, and used the more efficient transfection method, nucleofection, to deliver a construct of minimalistic, immunologically defined gene expression encoding the erythropoietin (MIDGE-EPO) into hMSC. MIDGE construct is relatively safer than the viral and plasmid expression systems as the detrimental eukaryotic and prokaryotic gene and sequences have been eliminated. Using a plasmid encoding the luciferase gene, we demonstrated a high transfection efficiency using the U-23 (21.79 � 1.09%) and C-17 (5.62 � 1.09%) pulsing program in nucleofection. The cell viabilities were (44.93 � 10.10)% and (21.93 � 5.72)%, respectively 24 h post-nucleofection. On the other hand, lipofection treatment only yielded less than 0.6% efficiencies despite showing higher viabilities. Nucleofection did not affect hMSC renewability, immunophenotype and differentiation potentials. Subsequently, we nucleofected MIDGE-EPO using the U-23 pulsing program into hMSC. The results showed that, despite a low nucleofection efficiency with this construct, the EPO protein was stably expressed in the nucleofected cells up to 55 days when determined by ELISA or immunocytochemical staining. In conclusion, nucleofection is an efficient non-viral transfection approach for hMSC, which when used in conjunction with a MIDGE construct, could result in extended and stable transgene expression in hMSC. � 2011 Springer Science+Business Media B.V.en_US
dc.description.natureFinalen_US
dc.identifier.CODENCYTOE
dc.identifier.doi10.1007/s10616-011-9413-2
dc.identifier.epage216
dc.identifier.issn9209069
dc.identifier.issue2
dc.identifier.scopus2-s2.0-84859157173
dc.identifier.spage203
dc.identifier.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-84859157173&doi=10.1007%2fs10616-011-9413-2&partnerID=40&md5=560de03dddd7de9120e58779fd339849
dc.identifier.urihttps://oarep.usim.edu.my/handle/123456789/10082
dc.identifier.volume64
dc.languageEnglish
dc.language.isoen_USen_US
dc.relation.ispartofCytotechnology
dc.sourceScopus
dc.subjectBone marrow mesenchymal stromal cellsen_US
dc.subjectCationic lipofectionen_US
dc.subjectErythropoietinen_US
dc.subjectMIDGEen_US
dc.subjectNucleofectionen_US
dc.titleExtended and stable gene expression via nucleofection of MIDGE construct into adult human marrow mesenchymal stromal cellsen_US
dc.title.alternativeCytotechnologyen_US
dc.typeArticleen_US
dspace.entity.typePublication

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