Kryokonservierung von hepatischen Sternzellen der Ratte

In this work a protocoll for the cryopreservation of rat hepatic stellate cells was developed. Isolated HSC of the rat are used as a model of liver fibrogenesis. Many proceedings are studied on this cells, for example the biotransformation, the gene expression, the effect of pharmaceuticals, different kinds of transcriptional factors and many more. Methods for the isolation of rat hepatic stellate cells and their cultivation are well established since years. Since today the discussion about the origin of HSC and different HSC subpopulations is not finished, and there is still room for speculations.The classical theory describes the septum trasnversum as origin of the hepatic stellate cells, in contrast a newer theory describes a neuroectodermal origin.1 In addition to this some examinations showed that isolated HSC are abel to express different kinds of proteins in different quantity (as example different kinds of collagene and alpha-smooth-muscle-actin).7 Because of this heterogeneity it could be very interesting to change isolated HSC between different laboratories. The cryopreservation and the exchange on dry ice could be a practical solution. Therefore a successful proceeding for the longtime storage of HSC was developed. Many parameters has been examined for example the freezing rates, the prefreeze processing the different cryoprotectants, the thawing conditions, the cell account by ml freezing solution and the storage conditions. The vitality, the cell morphology and the cell characteristic has been examined with untouched HSC by many different tests. Light and electronic microscopic analysis, cell proliferation assays, apoptosis testing, Oil Red O-staining, SDS-Page and immunoblotting of different marker proteins for fibrogenesis has been passed through. These examinations showed no significant differences between the cryopreserved and untouched cells. Neither the cell morphology nor the biochemical and cellular qualities are impaired by the cryopreservation process. The storage of cryopreserved HSC showed that a longtime storage of HSC between one till six months is possible with the devoleped protocol. These results showed us that cryopreserved HSC are untouched in their functionality. Consequently the cryopreservation is a practical possibility for the conservation and the exchange of HSC between laboratories.

Grantor dc:publisher

Publikationsserver der RWTH Aachen University

Year dc:date

2007

Author dc:creator

• Neyzen, Svenja

Contributors dc:contributor

• Weiskirchen, Ralf

• info:eu-repo/classification/ddc/610
• Kryokonservierung
• Leberfibrose
• Apoptosis
• Immunoblot
• Medizin
• Hepatische Sternzellen
• p 53
• CRP2
• Elektronenmikroskopie
• DMSO
• hepatic stellate cells
• Oil-Red-O-Staining

Statement dc:rights

• info:eu-repo/semantics/openAccess

Language dc:language

ger

Identifier

https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-114600%22

OAI record oai:identifier

https://publications.rwth-aachen.de/oai2d?verb=GetRecord&metadataPrefix=oai_dc&identifier=oai%3Apublications.rwth-aachen.de%3A52371