Share

Cryopreservation of keratinocytes in a monolayer.

Author(s) : PASCH J., SCHIEFER A., HESCHEL I., et al.

Type of article: Article

Summary

Cryopreservation of cells in tissues is a major challenge in cryobiology, especially in tissue engineering. It is very questionable whether protocols developed for the cryopreservation of isolated cells are also applicable for cells in more complex structures such as tissues. The aim of this study was to find an optimum cryopreservation protocol for keratinocytes in a monolayer (two-dimensional structure). These epidermal cells can be transplanted as a monolayer grown on an appropriate matrix for the treatment of deep-dermal burns and leg ulcers. Successful cryopreservation of such transplants would offer the advantage of long-term storage and immediate availability. In the paper, the variables investigated were the cryoprotective solution and the cooling rate. In order to find a nontoxic cryoprotective agent (CPA) which could be transplanted without additional washing, the authors concluded hydroxyethyl starch (HES) as a possible CPA in their experimental protocol with the commonly used CPAs DMSO, glycerol, and ethylene glycol. The cell survival rate was determined by dye exclusion (trypan blue) and the cell metabolism was investigated by cell activity assay (alamarBlue). The cryopreservation protocol with 10 wt.-% HES gave the highest survival rate (72%) and also the highest metabolic activity of cells after thawing comparable values for the other CPAs were: DMSO, 48%; glycerol, 8%; and ethylene glycol, 10%.

Details

  • Original title: Cryopreservation of keratinocytes in a monolayer.
  • Record ID : 2001-0547
  • Languages: English
  • Source: Cryobiology - vol. 39 - n. 2
  • Publication date: 1999/09

Links


See other articles in this issue (4)
See the source

Indexing