Lab's home page: http://www.koki.hu/cdnb/

The goal of the team is to understand processes underlying the formation of the neuronal phenotype. For this end, neural stem cell clones had been established from brain vesicles of p53-deficient (NE-4C, NE-7C2) and wild type (WNE) mouse embryos. Pan-neurogenic processes have been investigated in the course of in vitro induced neurogenesis. Questions are addressed to i) the role of contact cell to cell interactions and signalling through adhesion molecules; ii) the expression of different pro-neural and neural genes; iii) the mechanisms behind the determination of the neurotransmitter-phenotype; iv) the potential role of defined positional genes in the tuning of the neuronal phenotype. Using co-culture systems and labelled sub-clones of neural stem cells, the role of astrocytes on neuronal cell fate decision has been also addressed. By implanting labelled stem cells, the team searches for stem cell features and conditions, which could help the integration of stem cells into the brain tissue. In order to understand stem cell "homing", several methods were established or are under development in order to study the adhesive behaviour of stem cells.

The Laboratory of Cellular and Developmental Neurobiology was founded in January 1997. The staff members, some former associate and assistant professors of Eötvös Loránd University are involved in higher education and keep on working as part-time professors. Since the formation, the unit has been studying the mechanisms of neuron formation both in vitro by using neurogenic models of neural stem cells and in vivo by grafting stem cells into the brain of healthy and disease-model animals. The team is skilled in cell- and molecular biological techniques, as well as in surgical and histological methods. Most of their studies have been carried out on the pheno- and genotypically identical, NE-4C and NE-7C2 neuroectodermal progenitors cloned from the anterior brain vesicles of 9-day old, p53^-/- mouse embryos. These cells divide continuously in "maintaining cultures" and display neural stem cell features. In the presence of all-trans-retionic acid (RA) or perinatal astrocytes, they differentiate into neurons (in 5-7days) and later (~10 days) to astrocytes. In vitro neuron formation proceeds through well-defined stages with specific morphological, molecular and physiological characteristics. The cell clones represent immortalised models of embryonic neural stem cells. Recently, the team has cloned similar progenitors (WNE) from wild type mouse embryos. Labelled sub-clones of NE-4C cells expressing green fluorescent protein (GFP) or heat-resistant placental alkaline phosphatase (PLAP) were established. Deposition of the clones into ATCC is in progress.

 

For investigating "stem cell homing", they invented special assays, among them optical waveguide light spectroscopic (OWLS) assay techniques for quantifying cell-ECM adhesivity. They work in close collaboration with the manufacturer of OWLS instrumentation (MicroVacuum Ltd, Hungary) in development of OWLS assays for cell biological use. The cell motility and migratory patterns of developing neural cells have been investigated by computer-aided time-lapse cinematography, in close collaboration with the Dept. of Biological Physics, Eötvös L. Univ.

 

Labelled and non-labelled NE-4C cells have been used in a number of laboratories in frames of multiple collaborations. By producing in vitro chimera embryos, the developmental potential of early embryonic neural stem cells has been investigated in the Laboratory of Embryology, Agricultural Research Centre, Gödöllő (Dr. E. Gócza). Changes of gene expression in the course of in vitro induced neurogenesis have been investigated in Dr. J.Beckers laboratory (GSF, Neuherberg, Germany). The intracerebral fate of the cells had been investigated in collaboration with  Dr.T. Jaffredo (Inst.of Embryiology College de France, Nogent-sur Marne) and  Dr. J.P. Herman ( UMR-6544 ICNE, CNRS-Univ. de l'Mediterranée, Marseille, France).

 

Besides their own research, the Laboratory provides tissue culture facilities for all units of IEM-HAS and provides regular theoretical (1 semester) and technical (48 hrs) "In vitro Cell Technology" courses for Ph.D. students. The team gives a regular, 2-semester Ph.D. course on "Neural cell differentiation".