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Article: A 3D collagen microsphere culture system for GDNF-secreting HEK293 cells with enhanced protein productivity

TitleA 3D collagen microsphere culture system for GDNF-secreting HEK293 cells with enhanced protein productivity
Authors
Keywords3D culture
Biopharmaceutical manufacturing
Collagen
GDNF-secreting HEK293 cells
Microencapsulation
Microsphere
Issue Date2007
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/biomaterials
Citation
Biomaterials, 2007, v. 28 n. 35, p. 5369-5380 How to Cite?
Abstract
Mammalian cell culture technology has been used for decades in mass production of therapeutic proteins. However, unrestricted cell proliferation usually results in low-protein productivity. Controlled proliferation technologies such as metabolism intervention and genetic manipulation are therefore applied to enhance the productivity. Nevertheless, these strategies induced growth arrest with reduced viability and increased apoptosis. In this study, we report a new controlled proliferation technology by encapsulating human embryonic kidney (HEK) 293 cells over-expressing glial-derived neurotrophic factor (GDNF) in 3D collagen microspheres for extended culture. We investigated the viability, proliferation, cell cycle and GDNF productivity of HEK293 cells in microspheres as compared to monolayer culture. This system provides a physiologically relevant tissue-like environment for cells to grow and exerts proliferation control throughout the culture period without compromising the viability. A significant increase in the production rate of GDNF was found in the 3D microsphere system comparing with the monolayer culture. GDNF productivity was also significantly affected by the initial cell number and the serum concentration. The secreted GDNF was still bioactive as it induced neurite extension in PC12 cells. In summary, the 3D collagen microsphere system presents a cost-effective controlled growth technology for protein production in pharmaceutical manufacturing. © 2007 Elsevier Ltd. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/68318
ISSN
2013 Impact Factor: 8.312
ISI Accession Number ID
References

 

Author Affiliations
  1. The University of Hong Kong Li Ka Shing Faculty of Medicine
  2. Faculty of Engineering
DC FieldValueLanguage
dc.contributor.authorWong, HLen_HK
dc.contributor.authorWang, MXen_HK
dc.contributor.authorCheung, PTen_HK
dc.contributor.authorYao, KMen_HK
dc.contributor.authorChan, BPen_HK
dc.date.accessioned2010-09-06T06:03:27Z-
dc.date.available2010-09-06T06:03:27Z-
dc.date.issued2007en_HK
dc.identifier.citationBiomaterials, 2007, v. 28 n. 35, p. 5369-5380en_HK
dc.identifier.issn0142-9612en_HK
dc.identifier.urihttp://hdl.handle.net/10722/68318-
dc.description.abstractMammalian cell culture technology has been used for decades in mass production of therapeutic proteins. However, unrestricted cell proliferation usually results in low-protein productivity. Controlled proliferation technologies such as metabolism intervention and genetic manipulation are therefore applied to enhance the productivity. Nevertheless, these strategies induced growth arrest with reduced viability and increased apoptosis. In this study, we report a new controlled proliferation technology by encapsulating human embryonic kidney (HEK) 293 cells over-expressing glial-derived neurotrophic factor (GDNF) in 3D collagen microspheres for extended culture. We investigated the viability, proliferation, cell cycle and GDNF productivity of HEK293 cells in microspheres as compared to monolayer culture. This system provides a physiologically relevant tissue-like environment for cells to grow and exerts proliferation control throughout the culture period without compromising the viability. A significant increase in the production rate of GDNF was found in the 3D microsphere system comparing with the monolayer culture. GDNF productivity was also significantly affected by the initial cell number and the serum concentration. The secreted GDNF was still bioactive as it induced neurite extension in PC12 cells. In summary, the 3D collagen microsphere system presents a cost-effective controlled growth technology for protein production in pharmaceutical manufacturing. © 2007 Elsevier Ltd. All rights reserved.en_HK
dc.languageengen_HK
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/biomaterialsen_HK
dc.relation.ispartofBiomaterialsen_HK
dc.rightsBiomaterials. Copyright © Elsevier BV.en_HK
dc.subject3D cultureen_HK
dc.subjectBiopharmaceutical manufacturingen_HK
dc.subjectCollagenen_HK
dc.subjectGDNF-secreting HEK293 cellsen_HK
dc.subjectMicroencapsulationen_HK
dc.subjectMicrosphereen_HK
dc.subject.meshAnimalsen_HK
dc.subject.meshCell Culture Techniques - methodsen_HK
dc.subject.meshCell Lineen_HK
dc.subject.meshCollagen Type Ien_HK
dc.subject.meshGlial Cell Line-Derived Neurotrophic Factor - biosynthesis - secretionen_HK
dc.subject.meshHumansen_HK
dc.subject.meshKidney - cytology - secretionen_HK
dc.subject.meshMiceen_HK
dc.subject.meshMicrospheresen_HK
dc.subject.meshPC12 Cellsen_HK
dc.subject.meshRatsen_HK
dc.subject.meshRecombinant Proteins - biosynthesis - secretionen_HK
dc.titleA 3D collagen microsphere culture system for GDNF-secreting HEK293 cells with enhanced protein productivityen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0142-9612&volume=28&spage=5369&epage=5380&date=2007&atitle=A+3D+collagen+microsphere+culture+system+for+GDNF-secreting+HEK293+cells+with+enhanced+protein+productivityen_HK
dc.identifier.emailCheung, PT:ptcheung@hkucc.hku.hken_HK
dc.identifier.emailYao, KM:kmyao@hku.hken_HK
dc.identifier.emailChan, BP:bpchan@hkucc.hku.hken_HK
dc.identifier.authorityCheung, PT=rp00351en_HK
dc.identifier.authorityYao, KM=rp00344en_HK
dc.identifier.authorityChan, BP=rp00087en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.biomaterials.2007.08.016en_HK
dc.identifier.pmid17764735en_HK
dc.identifier.scopuseid_2-s2.0-34948825863en_HK
dc.identifier.hkuros145968en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-34948825863&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume28en_HK
dc.identifier.issue35en_HK
dc.identifier.spage5369en_HK
dc.identifier.epage5380en_HK
dc.identifier.eissn1878-5905-
dc.identifier.isiWOS:000250860000016-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridWong, HL=22236077400en_HK
dc.identifier.scopusauthoridWang, MX=22236053000en_HK
dc.identifier.scopusauthoridCheung, PT=7202595465en_HK
dc.identifier.scopusauthoridYao, KM=7403234578en_HK
dc.identifier.scopusauthoridChan, BP=7201530390en_HK

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