title: In vitro culture and oxygen consumption of NSCs in size-controlled neurospheres of Ca-alginate/gelatin microbead.
abstract: Neural stem cells (NSCs) forming neurospheres in a conventional culture tend to develop necrotic/apoptotic centers due to mass transport limitations. In this study, the internal pore structure of calcium-alginate/gelatin (CAG) microbeads was tuned and controlled to provide a suitable three-dimensional environment supporting NSC proliferation. Direct impact of three-dimensional space availability was quantified by oxygen consumption rates of NSCs and cells were cultured in three different methods: neurospheres, single cell suspension of NSCs, and encapsulated NSCs in microbeads. Our results showed that encapsulated NSCs in CAG microbeads maintained higher cell viability than in conventional culture. In addition, NSCs encapsulated in CAG microbeads preserved their original stemness and continued to express nestin, CNPase, GFAP and β-tubulin-III post-encapsulation. Oxygen consumption rates of encapsulated NSCs in CAG microbeads were the lowest as compared to the other two culture methods. The optimal cell density supporting high cell proliferation in CAG microbeads was found to be 1.5×10(5)cells/mL. The glucose consumption curve suggests that encapsulated NSCs in microbeads had a slower growth profile. This study presents an alternative method in hybrid microbead preparation to generate a highly favorable three-dimensional cell carrier for NSCs and was successfully applied for its effective in vitro expansion.
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China. Electronic address: Kedongsong@dlut.edu.cn.
authors.forename: Kedong
authors.initials: K
authors.lastname: Song
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China.
authors.forename: Yanfei
authors.initials: Y
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China.
authors.forename: Shixiao
authors.initials: S
authors.lastname: Li
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China.
authors.forename: Meiling
authors.initials: M
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China.
authors.forename: Yixing
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore, Singapore.
authors.forename: Mayasari
authors.initials: M
authors.lastname: Lim
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China. Electronic address: liutq@dlut.edu.cn.
authors.forename: Tianqing
authors.initials: T
authors.lastname: Liu
mesh_terms: D000464:Alginates / Q000737:chemistry*; D000818:Animals; D002118:Calcium / Q000737:chemistry*; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D005780:Gelatin / Q000737:chemistry*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D051379:Mice; D008863:Microspheres; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D010101:Oxygen Consumption
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D005780:Gelatin; D002118:Calcium
keywords: Calcium-alginate/gelatin microbeads; Encapsulation; Neural stem cells; Oxygen consumption; Size control