Development of Systematic Parameter Optimization for Cryopreservation Protocols for Cellular Suspensions

Authors

  • N. Hofmann Leibniz Universitaet Hannover
  • I. Bernemann Leibniz Universitaet Hannover
  • D. Pogozhykh Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine
  • B. Glasmacher Leibniz Universitaet Hannover

Keywords:

cryopreservation, cell suspensions, human keratinocytes, HPMEC-ST1.6R, cryoprotectants, Me2SO, cooling rate

Abstract

Effective cryopreservation of cellular suspensions is an important task of cryobiology. A systematic parameter optimization setup for cryopreservation of cellular suspensions was established on the basis of our previous studies. Keratinocytes were used as a cellular model. The investigation included 48 protocols with different cooling rates (B) in a two-step freezing protocols (B1 in the range of 4°C to –30°C and B2 from –30°C to –80°C). All protocols have been studied with four different concentrations of the most frequently applied cryoprotective agent dimethyl sulfoxide (DMSO). Raster analysis of approximately 600 samples allowed the detection of the most comprehensive array of possible variables. In further investigations this setup was used to improve the cryopreservation protocols for HPMEC-ST1.6R (human pulmonary microvascular endothelial cells). Results revealed that only an optimal combination of cooling rates ensured the highest survival rates of cells after thawing. In case of keratinocytes, the optimal freezing protocol was composed of the cooling rates of 5 K/min for B1 and 7.5 K/min for B2, or 5 K/min for B1 and 10 K/min for B2 (2.5% DMSO concentration). The highest survival rates of HPMEC-ST1.6R cell culture were achieved with application of the following protocols: 5 K/min (B1) combined with 5 K/min (B2) (7.5% DMSO); 3 K/min, combined with 3 K/min (5% or 7.5% DMSO); and 10 K/min combined with 3 K/min (10% DMSO).

Downloads

Published

2011-12-19

How to Cite

Hofmann, N., Bernemann, I., Pogozhykh, D., & Glasmacher, B. (2011). Development of Systematic Parameter Optimization for Cryopreservation Protocols for Cellular Suspensions. Problems of Cryobiology and Cryomedicine, 21(4), 353–364. Retrieved from https://cryo.org.ua/journal/index.php/probl-cryobiol-cryomed/article/view/107

Issue

Vol. 21 No. 4 (2011): Problems of Cryobiology

Section

Theoretical and Experimental Cryobiology

License

Copyright (c) 2020 N. Hofmann, I. Bernemann, D. Pogozhykh, B. Glasmacher

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Authors who publish with this journal agree to the following terms:

  • Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
  • Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
  • Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).