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A comparative study of freezing single cells and spheroids: Towards a new model system for optimizing freezing protocols for cryobanking of human tumours

: Ehrhart, F.; Schulz, J.C.; Katsen-Globa, A.; Shirley, S.G.; Reuter, D.; Bach, F; Zimmermann, U.; Zimmermann, H.


Cryobiology 58 (2009), No.2, pp.119-127
ISSN: 0011-2240
ISSN: 1090-2392
Journal Article
Fraunhofer IBMT ()

Cryopreservation of human turnout cells and tissue is a valuable tool for retrospective analysis and for the transport and handling of biopsy material. Tumour tissue consists of different cell types, which have different optimal freezing conditions, and extracellular matrix. A well-defined and authentic model system is required for developing new freezing protocols and media. This work describes the use of 1.929 and PC-3 spheroids as new model systems for freezing human tumours. Cell suspension and spheroids were frozen in different vessels (1 ml cryovials and a special, cryo-compatible 30 x 25 mu l multi well plate) at slow rate (1 degrees C/min). Freezing media were combinations of culture or tumour transport medium (Liforlab (R)) with the cryoprotective agents, Me2SO, trehalose and modified starch. We also present a new method of evaluating the viability of three dimensional multicellular systems to compare thawed spheroids objectively. Best viability (70%) of L929 spheroids occurred with a combination of Liforlab (R) and starch hydrolysis product. The best cryopreservation results for spheroids were found with extracellular cryoprotectants, while optimum viability of single cells was achieved with Me2SO.