title: Enhanced survival and neurite network formation of human umbilical cord blood neuronal progenitors in three-dimensional collagen constructs.
abstract: Umbilical cord blood (CB) stem cells have been proposed for cell-based therapeutic applications for diverse diseases of the CNS. We hypothesized that tissue-engineering strategies may extend the efficacy of these approaches by improving the long-term viability and function of stem cell-derived neuronal progenitors. To test our hypothesis, we explored the survival and differentiation of human CB-derived neuronal progenitors (HUCBNP) in a three-dimensional (3D) collagen construct. In contrast to two-dimensional culture conditions, the cells survived in 3D for an extended period of time of more than 2 months. Under 3D conditions, HUCBNP underwent spontaneous neuronal differentiation, which was further enhanced by treatment with neuronal conditioned medium (CM) and nerve growth factor (NGF). Neurite outgrowth, quantified by assessing the fractal dimension (D f) of the complex neuronal networks, was significantly enhanced under 3D conditions in the presence of CM/NGF, concomitant with a reduced expression of the early neuronal marker nestin (1.9-fold), and increased levels of mature neuronal markers such as MAP-2 (3.6-fold), β-tubulin (1.5-fold), and neuronal specific enolase (6.6-fold) and the appearance of the synaptic marker synaptophysin. To assess the feasibility for clinical usage, HUCBNP were also isolated from frozen CB samples and cultured under 3D conditions. The data indicate the essential complete preservation of neurotrophic (survival) and neurotropic (neurite outgrowth) properties. In conclusion, 3D culture conditions are proposed as an essential step for both maintenance of CB neuronal progenitors in vitro and for investigating specific features of neuronal differentiation towards future use in regenerative therapy.
authors.affiliation: School of Pharmacy, Institute for Drug Research, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, 91120, Israel.
authors.forename: Marian M
authors.initials: MM
authors.lastname: Bercu
authors.forename: Hadar
authors.initials: H
authors.lastname: Arien-Zakay
authors.forename: Dana
authors.initials: D
authors.lastname: Stoler
authors.forename: Shimon
authors.initials: S
authors.lastname: Lecht
authors.forename: Peter I
authors.initials: PI
authors.lastname: Lelkes
authors.forename: Simcha
authors.initials: S
authors.lastname: Samuel
authors.forename: Reuven
authors.initials: R
authors.lastname: Or
authors.forename: Arnon
authors.initials: A
authors.lastname: Nagler
authors.forename: Philip
authors.initials: P
authors.lastname: Lazarovici
authors.forename: Uriel
authors.initials: U
authors.lastname: Elchalal
mesh_terms: D002470:Cell Survival; D003094:Collagen / Q000494:pharmacology*; D015925:Cryopreservation; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D020932:Nerve Growth Factor / Q000494:pharmacology; D064231:Nestin / Q000235:genetics / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D055495:Neurogenesis; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000378:metabolism; D016708:Synaptophysin / Q000235:genetics / Q000378:metabolism; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry*; D014404:Tubulin / Q000235:genetics / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; C489788:MAP2 protein, human; D008869:Microtubule-Associated Proteins; D064231:Nestin; D016708:Synaptophysin; D014404:Tubulin; D003094:Collagen; D020932:Nerve Growth Factor; D010751:Phosphopyruvate Hydratase