title: Efficient animal-serum free 3D cultivation method for adult human neural crest-derived stem cell therapeutics.
abstract: Due to their broad differentiation potential and their persistence into adulthood, human neural crest-derived stem cells (NCSCs) harbour great potential for autologous cellular therapies, which include the treatment of neurodegenerative diseases and replacement of complex tissues containing various cell types, as in the case of musculoskeletal injuries. The use of serum-free approaches often results in insufficient proliferation of stem cells and foetal calf serum implicates the use of xenogenic medium components. Thus, there is much need for alternative cultivation strategies. In this study we describe for the first time a novel, human blood plasma based semi-solid medium for cultivation of human NCSCs. We cultivated human neural crest-derived inferior turbinate stem cells (ITSCs) within a blood plasma matrix, where they revealed higher proliferation rates compared to a standard serum-free approach. Three-dimensionality of the matrix was investigated using helium ion microscopy. ITSCs grew within the matrix as revealed by laser scanning microscopy. Genetic stability and maintenance of stemness characteristics were assured in 3D cultivated ITSCs, as demonstrated by unchanged expression profile and the capability for self-renewal. ITSCs pre-cultivated in the 3D matrix differentiated efficiently into ectodermal and mesodermal cell types, particularly including osteogenic cell types. Furthermore, ITSCs cultivated as described here could be easily infected with lentiviruses directly in substrate for potential tracing or gene therapeutic approaches. Taken together, the use of human blood plasma as an additive for a completely defined medium points towards a personalisable and autologous cultivation of human neural crest-derived stem cells under clinical grade conditions.
authors.affiliation: Molecular Neurobiology, University of Bielefeld, Universitaetsstr. 25, D-33501 Bielefeld, Germany.
authors.forename: J F W
authors.initials: JF
authors.lastname: Greiner
authors.forename: S
authors.initials: S
authors.lastname: Hauser
authors.forename: D
authors.initials: D
authors.lastname: Widera
authors.forename: J
authors.initials: J
authors.lastname: Müller
authors.forename: F
authors.initials: F
authors.lastname: Qunneis
authors.forename: C
authors.initials: C
authors.lastname: Zander
authors.forename: I
authors.initials: I
authors.lastname: Martin
authors.forename: J
authors.initials: J
authors.lastname: Mallah
authors.forename: D
authors.initials: D
authors.lastname: Schuetzmann
authors.forename: C
authors.initials: C
authors.lastname: Prante
authors.forename: H
authors.initials: H
authors.lastname: Schwarze
authors.forename: W
authors.initials: W
authors.lastname: Prohaska
authors.forename: A
authors.initials: A
authors.lastname: Beyer
authors.forename: K
authors.initials: K
authors.lastname: Rott
authors.forename: A
authors.initials: A
authors.lastname: Hütten
authors.forename: A
authors.initials: A
authors.lastname: Gölzhäuser
authors.forename: H
authors.initials: H
authors.lastname: Sudhoff
authors.forename: C
authors.initials: C
authors.lastname: Kaltschmidt
authors.forename: B
authors.initials: B
authors.lastname: Kaltschmidt
mesh_terms: D000943:Antigens, Differentiation / Q000378:metabolism; D040761:Biomimetic Materials; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D016895:Culture Media, Serum-Free; D005337:Fibrin / Q000648:ultrastructure; D020869:Gene Expression Profiling; D006801:Humans; D057139:Nanofibers / Q000648:ultrastructure; D009416:Nerve Regeneration; D009432:Neural Crest / Q000166:cytology*; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D010949:Plasma; D016062:Porosity; D018874:Spheroids, Cellular / Q000166:cytology
chemical_list: D000943:Antigens, Differentiation; D016895:Culture Media, Serum-Free; D005337:Fibrin