Magnetic-Activated-Cell-Sorting (MACS); Entering guide in method - a possibility to separate fetal neural rat stem cells expressing the GABAA-receptor gamma 2 from different types of lineage-restricted-porgenitors
In the first part of the dissertation the method of magnetic cell sorting was opened up and proved for practice. Gamma-aminobutyric-acid is the principal inhibitory neurotransmitter in the brain. Apart from the important role in the synaptic transmission, gamma-aminobutyric-acid also provides signalling indices in the developing nervous system. A positive effect on the labeling steps by using buffer from the companies could neither been shown nor excluded. Furthermore it was shown that a combination of the pasteur principle plus sample separation fi lters is an excellent method to get a single cell suspension necessary for correct labeling. It could also be helpful to reprocess older cell populations. Furthermore there were indices that cells treated with magnetic cell sorting have a higher mortality rate than original population in cell culture. As the gamma-aminobutyric-acid-A receptor is crurically participating in regulation of differentiation and proliferation of neural progenitor cells we aimed to isolate gamma-aminobutyric-acid-receptor positive stem cells. Therefore the treated cells were documented by immunostaining. Only in long term differentiation we observed a higher expression of gammaaminobutyric-acid-receptor in the labeled fraction than in the unlabeled fraction. Admittedly the labeled fraction seem to show a different nestin expression than the unlabeled fraction after forteen days of differentiation, but a higher nestin exprimation in the gamma-aminobutyric-acidreceptor positive fraction which points at a higher stem cell rate in the labeled fraction, could not be shown. Consequently, a separation of fetal neural rat stem cells expressing the gamma-aminobutyric-acid-A receptor gamma 2 from different types of LRPs with the method called magnetic activated cell sorting could not be established with the antigens used in dissertation.
Erstellung / Fertigstellung
Normierte SchlagwörterMagnetaktivierter Zellsortierer [GND]
Flow cytometry [MeSH]
Neural stem cells [MeSH]
Receptors, GABA-A [MeSH]