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Impressum
Morphological Differentiation of Adult Human Mesenchymal Progenitor Cells into Cells with a Neuronal Phenotype in vitro
1Linke S., 2Stute N., 2Zander A., 1Richard G., 3Schachner M., 3Bartsch U.
1Augenklinik und 2Einrichtung für Knochenmarktransplantation, Universitätsklinikum Eppendorf, Martinistr. 52, 20 246 Hamburg; 3Institut für Biosynthese neuraler Strukturen; Zentralinstitut für Molekulare Neurobiologie Hamburg (ZMNH)
Objective: To determine the differentiation potential of adult
human mesenchymal progenitor cells (AI Caplan`s method of isolation and
cultivation) into nerve cells in vitro.
Methods: Adult human mesenchymal progenitor cells (MPC`s) were
harvested from volunteers with informed consent and expanded in culture
as described previously (Haynesworth et al. 1992). In short, bone marrow
mononuclear cells were separated by ficoll-gradient centrifugation and
transferred to plastic culture dishes containing DMEM-medium and 10% fetal
bovine serum (FBS, selected lot). After the third passage these mesenchymal
progenitor cells were plated at a density of 2000 cells/cm² on PLL-coated
coverslips and induced to differentiate by removal of FBS and addition
of 2%DMSO/200µM butylated hydroxianisole (Woodbury et al. 2000).
Morphological differentiation was analyzed by phase contrast microscopy
and time lapse video-microscopy.
Results: Adult human mesenchymal progenitor cells could be rapidly
expanded in our culture conditions. After three passages, this undifferentiated
and homogenous cell population (confirmed by FACSanalysis) could be induced
to differentiate into cells displaying a neuronal morphology. Within 45
minutes after initiation of differentiation, some of the spindle-shaped
MPC`s became spherical, round-shaped and subsequently extended neurite-like
processes. Most of the cells differentiated into cells with a bipolar
morphology within several hours. After an eight-hour incubation time in
differentiation medium more than 75% of the cells morphologically displayed
a neuronal phenotype.
Conclusions: Undifferentiated, non hematopoietic cells derived
from adult human bone marrow could be induced to rapidly differentiate
into cells displaying a neuron-like morphology. Experiments are in progress
to further characterize these cells immunocytochemically and to determine
the fate of these cells after transplantation into the pathologically
altered mouse retina.