title: Insights in spatio-temporal characterization of human fetal neural stem cells.
abstract: Primary human fetal cells have been used in clinical trials of cell replacement therapy for the treatment of neurodegenerative disorders such as Huntington's disease (HD). However, human fetal primary cells are scarce and difficult to work with and so a renewable source of cells is sought. Human fetal neural stem cells (hfNSCs) can be generated from human fetal tissue, but little is known about the differences between hfNSCs obtained from different developmental stages and brain areas. In the present work we characterized hfNSCs, grown as neurospheres, obtained from three developmental stages: 4-5, 6-7 and 8-9weeks post conception (wpc) and four brain areas: forebrain, cortex, whole ganglionic eminence (WGE) and cerebellum. We observed that, as fetal brain development proceeds, the number of neural precursors is diminished and post-mitotic cells are increased. In turn, primary cells obtained from older embryos are more sensitive to the dissociation process, their viability is diminished and they present lower proliferation ratios compared to younger embryos. However, independently of the developmental stage of derivation proliferation ratios were very low in all cases. Improvements in the expansion rates were achieved by mechanical, instead of enzymatic, dissociation of neurospheres but not by changes in the seeding densities. Regardless of the developmental stage, neurosphere cultures presented large variability in the viability and proliferation rates during the initial 3-4 passages, but stabilized achieving significant expansion rates at passage 5 to 6. This was true also for all brain regions except cerebellar derived cultures that did not expand. Interestingly, the brain region of hfNSC derivation influences the expansion potential, being forebrain, cortex and WGE derived cells the most expandable compared to cerebellar. Short term expansion partially compromised the regional identity of cortical but not WGE cultures. Nevertheless, both expanded cultures were multipotent and kept the ability to differentiate to region specific mature neuronal phenotypes.
authors.affiliation: Laboratory of Stem Cells and Regenerative Medicine, Department of Biomedical Science, Faculty of Medicine and Health Sciences, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; August Pi i Sunyer Biomedical Research Institute (IDIBAPS), Rosselló 149-153, 08036 Barcelona, Spain; Networked Biomedical Research Centre for NeuroDegenerative Disorders (CIBERNED), Spain; Research and Development Unit, Cell Therapy Program, Faculty of Medicine and Health Sciences, University of Barcelona, Casanova 143, 08036 Barcelona, Spain. Electronic address: r.martin@ub.edu.
authors.forename: Raquel
authors.initials: R
authors.lastname: Martín-Ibáñez
authors.affiliation: Laboratory of Stem Cells and Regenerative Medicine, Department of Biomedical Science, Faculty of Medicine and Health Sciences, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; August Pi i Sunyer Biomedical Research Institute (IDIBAPS), Rosselló 149-153, 08036 Barcelona, Spain; Networked Biomedical Research Centre for NeuroDegenerative Disorders (CIBERNED), Spain. Electronic address: ines.guardia@ub.edu.
authors.forename: Inés
authors.initials: I
authors.lastname: Guardia
authors.affiliation: Laboratory of Stem Cells and Regenerative Medicine, Department of Biomedical Science, Faculty of Medicine and Health Sciences, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; August Pi i Sunyer Biomedical Research Institute (IDIBAPS), Rosselló 149-153, 08036 Barcelona, Spain; Networked Biomedical Research Centre for NeuroDegenerative Disorders (CIBERNED), Spain. Electronic address: monica.pardo@ub.edu.
authors.forename: Mónica
authors.initials: M
authors.lastname: Pardo
authors.affiliation: Laboratory of Stem Cells and Regenerative Medicine, Department of Biomedical Science, Faculty of Medicine and Health Sciences, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; August Pi i Sunyer Biomedical Research Institute (IDIBAPS), Rosselló 149-153, 08036 Barcelona, Spain; Networked Biomedical Research Centre for NeuroDegenerative Disorders (CIBERNED), Spain; Research and Development Unit, Cell Therapy Program, Faculty of Medicine and Health Sciences, University of Barcelona, Casanova 143, 08036 Barcelona, Spain. Electronic address: cherranzsotoca@ub.edu.
authors.forename: Cristina
authors.initials: C
authors.lastname: Herranz
authors.affiliation: Cardiff University Brain Repair Group, Schools of Biosciences and Medicine, University of Cardiff, UK. Electronic address: zietlow@mpiib-berlin.mpg.de.
authors.forename: Rike
authors.initials: R
authors.lastname: Zietlow
authors.affiliation: Cardiff University Brain Repair Group, Schools of Biosciences and Medicine, University of Cardiff, UK. Electronic address: vinhnn@cardiff.ac.uk.
authors.forename: Ngoc-Nga
authors.initials: NN
authors.lastname: Vinh
authors.affiliation: Cardiff University Brain Repair Group, Schools of Biosciences and Medicine, University of Cardiff, UK. Electronic address: rosserae@cardiff.ac.uk.
authors.forename: Anne
authors.initials: A
authors.lastname: Rosser
authors.affiliation: Laboratory of Stem Cells and Regenerative Medicine, Department of Biomedical Science, Faculty of Medicine and Health Sciences, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; August Pi i Sunyer Biomedical Research Institute (IDIBAPS), Rosselló 149-153, 08036 Barcelona, Spain; Networked Biomedical Research Centre for NeuroDegenerative Disorders (CIBERNED), Spain; Research and Development Unit, Cell Therapy Program, Faculty of Medicine and Health Sciences, University of Barcelona, Casanova 143, 08036 Barcelona, Spain. Electronic address: jmcanals@ub.edu.
authors.forename: Josep M
authors.initials: JM
authors.lastname: Canals
mesh_terms: D000704:Analysis of Variance; D001921:Brain / Q000166:cytology / Q000196:embryology; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology*; D002470:Cell Survival; D002478:Cells, Cultured; D053686:Fetal Stem Cells / Q000166:cytology* / Q000502:physiology; D005333:Fetus; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D005865:Gestational Age; D006801:Humans; D019394:Ki-67 Antigen; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D019394:Ki-67 Antigen; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger
keywords: Cell replacement; Cerebellum; Cortex; Differentiation; Expansion; Forebrain; Ganglionic eminence; Human fetal neural stem cells; Huntington's disease; Regional identity