Rapid cryopreservation of five mammalian and one mosquito cell line at −80°C while attached to flasks in a serum free cryopreservative

Abstract

Cell culturing, and the requisite storage of cell lines at ultra-low temperatures, is used in most laboratories studying or using eukaryotic proteomics, genomics, microarray, and RNA technologies. In this study we have observed that A72(dog), CRFK(cat), NB324K(human), MCF7(human), WI38(human), and C636(mosquito) cells were effectively cryopreserved at -80°C while attached to the substratum of 25cm 2 tissue culture flasks. This was accomplished using a serum free crypreservative recently developed by Corsini and co-workers. The technique allows for significant savings of time and money in laboratories that rapidly process numerous cell lines.

Directional freezing for the cryopreservation of adherent mammalian cells on a substrate

“…We suggest that the cooling rate in cell culture cryopreservation should be slowed down with the decrease of temperature, which is also in line with a protocol applied in embryo cryopreservation [ 36 ]. Our approach displays high reproducibility and high survival rates compared to non-directional slow freezing methods and other previously reported attempts of cryopreservation of cells in an adherent state [ 3 , 12 – 18 , 21 ]. Our method can be readily applied as a robust cryopreservation protocol for various in vitro applications.…”

“…Several studies have demonstrated the feasibility of cryopreserving cells adhered to a substrate [ 3 , 12 – 22 ] exploring the effects of CPAs composition, extracellular matrix design, and substrate modification. However, no in depth study has been reported on optimization of the cryopreservation procedure of adherent cultures in terms of cooling rates and ice geometry.…”

A Simple and Rapid Cryopreservation Technique for Ciliates: A Long‐Term Storage Procedure Used for Marine Scuticociliates

“…According to previous reports, various cell lines have been stored at −70 °C to −90 °C and successfully recovered (Bjornsson and Huebner ; Corsini and Mann ; Mazur ). Biochemical activities of ciliates are not totally stopped at such temperatures, and cellular damage and accumulation of toxic metabolites shorten the time for which cells remain viable during storage (Ashwood‐Smith and Farrant ; Gao and Critser ; Mazur ).…”