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55. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e. V. (DGNC)
1. Joint Meeting mit der Ungarischen Gesellschaft für Neurochirurgie

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

25. bis 28.04.2004, Köln

In vitro differentiation of expanded human fetal neuronal precursor cells

In-vitro-Differenzierung von menschlichen embryonalen neuronalen Vorläuferzellen

Meeting Abstract

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  • corresponding author Jaroslaw Maciaczyk - Labor für Molekulare Neurochirurgie, Abt. für Funktionelle und Stereotaktische Neurochirurgie, Universitätsklinikum Freiburg - Neurozentrum, Freiburg
  • G. Nikkhah - Labor für Molekulare Neurochirurgie, Abt. für Funktionelle und Stereotaktische Neurochirurgie, Universitätsklinikum Freiburg - Neurozentrum, Freiburg

Deutsche Gesellschaft für Neurochirurgie. Ungarische Gesellschaft für Neurochirurgie. 55. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC), 1. Joint Meeting mit der Ungarischen Gesellschaft für Neurochirurgie. Köln, 25.-28.04.2004. Düsseldorf, Köln: German Medical Science; 2004. DocP 03.28

The electronic version of this article is the complete one and can be found online at:

Published: April 23, 2004

© 2004 Maciaczyk et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.




Cell replacement therapy, based on human fetal cell transplantation, proved to be beneficial under experimental conditions and in clinical trials. The ethical concerns and limited availability of the aborted tissue make it difficult for this therapy to become a routine clinical strategy. One way to overcome these obstacles is the in vitro expansion of human fetal-derived neural precursor cells, their characterization and differentiation into the desired phenotype for further clinical applications.


Cortex (CTX) ganglionic eminences (STR), ventral midbrains (VM) and spinal cord (SC) were obtained from elective abortions up to 12th week post conception and expanded as a free floating aggregates (neurospheres) for prolonged periods of time in medium supplemented with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). For differentiation cells were dissociated, plated onto poly-L-lysine-laminin coated cover-slips and cultivated in mitogens deprived culture conditions for 14 days. Immunostaining against lineage specific markers - type III β-tubuline (TuJ1), microtubule associated protein 2 (MAP-2), glial fibrillary acid protein (GFAP), galactocerebroside (Gal-C) and nestin was performed. Neurons were characterized with several phenotype specific markers. The proliferative activity of the cultures was investigated with BrdU incorporation assay followed by short time of differentiation and immuocytochemical analysis.


All of the specimens expanded in vitro for more than 30 weeks with no signs of senescence. The majority of cells incorporated BrdU which was co-localized with both neuronal and glial markers. After differentiation precursors gave rise to neurons, astrocytes and oligodendrocytes. The expression of lineage specific markers was influenced by the origin of cultured tissue. The phylogenetically younger parts of the CNS (VM, SC) contained significant numbers of fully differentiated glial cells, whereas in cultures derived from telencephalon (CTX, STR) predominantly neurons with limited amounts of radial glia-like GFAP+ cells (CTX) or morphologically immature astrocytes (STR) were observed. The broad diversity of phenotype specific markers was detected in neurons from all cultured tissues with predominance of GAD+ cells (GABA-ergic neurons). Neurogranin- and ChAT- positive cells were more frequent in CTX and SC cultures but TH and serotonin reactivity was very limited.


The present study demonstrates the successful proliferation of human fetal-derived neural precursors and longterm differentiation into all cellular components of the CNS. The differentiation pattern was directly linked to the type of the tissue and age of the donor – resembling, in part, the ontogenetic maturation of the neuraxis. The in vitro proliferation of human fetal neural precursor cells might therefore be an alternative cell source for experimental and clinical research directed towards neural restorative strategies.