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Article: A 3D collagen microsphere culture system for GDNF-secreting HEK293 cells with enhanced protein productivity
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TitleA 3D collagen microsphere culture system for GDNF-secreting HEK293 cells with enhanced protein productivity
 
AuthorsWong, HL2
Wang, MX1
Cheung, PT1
Yao, KM1
Chan, BP2
 
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
 
CitationBiomaterials, 2007, v. 28 n. 35, p. 5369-5380 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.biomaterials.2007.08.016
 
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.
 
ISSN0142-9612
2013 Impact Factor: 8.312
 
DOIhttp://dx.doi.org/10.1016/j.biomaterials.2007.08.016
 
ISI Accession Number IDWOS:000250860000016
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorWong, HL
 
dc.contributor.authorWang, MX
 
dc.contributor.authorCheung, PT
 
dc.contributor.authorYao, KM
 
dc.contributor.authorChan, BP
 
dc.date.accessioned2010-09-06T06:03:27Z
 
dc.date.available2010-09-06T06:03:27Z
 
dc.date.issued2007
 
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.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationBiomaterials, 2007, v. 28 n. 35, p. 5369-5380 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.biomaterials.2007.08.016
 
dc.identifier.doihttp://dx.doi.org/10.1016/j.biomaterials.2007.08.016
 
dc.identifier.eissn1878-5905
 
dc.identifier.epage5380
 
dc.identifier.hkuros145968
 
dc.identifier.isiWOS:000250860000016
 
dc.identifier.issn0142-9612
2013 Impact Factor: 8.312
 
dc.identifier.issue35
 
dc.identifier.openurl
 
dc.identifier.pmid17764735
 
dc.identifier.scopuseid_2-s2.0-34948825863
 
dc.identifier.spage5369
 
dc.identifier.urihttp://hdl.handle.net/10722/68318
 
dc.identifier.volume28
 
dc.languageeng
 
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/biomaterials
 
dc.publisher.placeNetherlands
 
dc.relation.ispartofBiomaterials
 
dc.relation.referencesReferences in Scopus
 
dc.rightsBiomaterials. Copyright © Elsevier BV.
 
dc.subject.meshAnimals
 
dc.subject.meshCell Culture Techniques - methods
 
dc.subject.meshCell Line
 
dc.subject.meshCollagen Type I
 
dc.subject.meshGlial Cell Line-Derived Neurotrophic Factor - biosynthesis - secretion
 
dc.subject.meshHumans
 
dc.subject.meshKidney - cytology - secretion
 
dc.subject.meshMice
 
dc.subject.meshMicrospheres
 
dc.subject.meshPC12 Cells
 
dc.subject.meshRats
 
dc.subject.meshRecombinant Proteins - biosynthesis - secretion
 
dc.subject3D culture
 
dc.subjectBiopharmaceutical manufacturing
 
dc.subjectCollagen
 
dc.subjectGDNF-secreting HEK293 cells
 
dc.subjectMicroencapsulation
 
dc.subjectMicrosphere
 
dc.titleA 3D collagen microsphere culture system for GDNF-secreting HEK293 cells with enhanced protein productivity
 
dc.typeArticle
 
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Author Affiliations
  1. The University of Hong Kong Li Ka Shing Faculty of Medicine
  2. Faculty of Engineering