Verfahren zur Kryokonservierung epidermaler Zellen auf Kollagen-Microcarriern

The objective of this work was the development of a protocol for the cryopreservation of epidermal cells (keratinocytes) on collagen microcarriers. The cultivated carriers can be possibly used for the treatment of deep-dermal burns. The cryopreservation allows for the long-term storage of this transplant. The work is divided into the development of * a cryopreservation protocol for cells in a monolayer and * a modification and optimization of the collagen microcarriers with respect to the success of the cryopreservation and the cultivation after the thawing procedure. For the development of a cryopreservation protocol for the keratinocytes in a monolayer the cryoprotective agent (CPA) (DMSO, glycerol, ethylene glycol and HES) and the cooling rate (-1, -3 und -10°C/min) were modified. The best result was achieved with an HES concentration of 10 wt.-% and a cooling rate of -3°C/min. The obtained survival rate of the cells was 72%. No comparably satisfactory results were realized when using the other CPAs. In the second part of this work, the behavior of the microcarriers during the cryopreservation process was investigated on behalf of a newly designed cryo-microscope freezing stage. During the freezing phase a dramatic shrinkage of the microcarrier´s volume could be seen which led to the immediate detachment of the cells from the microcarrier. A similar shrinkage could be observed after exposure to a highly concentrated salt concentration at room temperature. By cross-linking the collagen in the shrunken state with glutardialdehyde the shrinkage during the freezing process could be significantly diminished. In addition, the seeding behaviour of the carrier with cells could be improved by an additional pretreament with soluble collagen. By combining the cryopreservation protocol (-3°C/min, 10 wt.-% HES) and the modification of the collagen microcarriers with salt and glutardialdehyde, the survival rate of the epidermal cells on the collagen microcarriers after cryopreservation could be improved from 3,3% to 38%.

Grantor dc:publisher

Publikationsserver der RWTH Aachen University

Year dc:date

2000

Author dc:creator

• Pasch, Julia

Contributors dc:contributor

• Rau, Günter

• info:eu-repo/classification/ddc/610
• Medizin

Statement dc:rights

• info:eu-repo/semantics/openAccess

Language dc:language

ger

Identifier

https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-118412%22

OAI record oai:identifier

https://publications.rwth-aachen.de/oai2d?verb=GetRecord&metadataPrefix=oai_dc&identifier=oai%3Apublications.rwth-aachen.de%3A56295