X-Message-Number: 15805
Date: Tue, 6 Mar 2001 03:47:31 -0800 (PST)
From: Doug Skrecky <>
Subject: Cryostor CS 5N maximises cryopreservation survival

Title
  Cell viability improves
  following inhibition of cryopreservation-induced apoptosis.
Source
  In Vitro Cellular & Developmental Biology. Animal. 
  36(4):262-70, 2000 Apr.
Abstract
  A new concept in cryopreservation solution design was developed that focuses
  on the use of an intracellular-type, hypothermic maintenance
  medium coupled with additives that inhibit cryopreservation-induced
  apoptosis. HypoThermosol' (HTS), a hypothermic (4 degrees C) maintenance
  medium utilized in the long-term storage of cell, tissue,
  and organ systems, was tested for cryoprotective capability on a renal
  cell line (Madin-Darby Canine Kidney
  cells). HTS and HTS derivatives were tested against
  conventional cell culture medium (Dulbecco's Minimal
  Essential medium, DME) as the cryoprotectant carrier solution because (1)
  cells are exposed to an extended state of hypothermia during
  the freeze-thaw process, and (2) HTS is designed to protect
  cells exposed to a hypothermic state. Cells
  separately cryopreserved in either HTS or DME + 5% dimethyl sulfoxide (DMSO)
  yielded equivalent 24-h postthaw survival (approximately 30%) and 5-d
  recovery (approximately 90%). Cells cryopreserved in
  CryoStor CS 5, a HTS derivative containing 5% DMSO, yielded approximately 75%
  24-h postthaw survival and recovery to 100% within 3 d. DNA gel
  electrophoresis was performed to determine the mechanisms of
  cell death contributing to cryopreservation failure.
  Cells preserved in DME (DMSO-free) died primarily through
  necrosis, whereas cells preserved in either DME + 5% DMSO,
  HTS, or CryoStor CS 5 died through a combination of apoptosis and necrosis.
  This observation led to the inclusion of an apoptotic inhibitor designed to
  improve cryopreservation outcome. MDCK cells cryopreserved
  in CryoStor CS 5 supplemented with an apoptotic inhibitor (Caspase I
  Inhibitor V), hereafter termed CryoStor CS 5N, resulted in a 24-h postthaw
  survival and recovery rate exceeding that of any other cryoprotective
  solution tested (85%). We conclude that: (1) the use of HTS (a dextran-based,
  intracellular-type solution) without DMSO can yield postthaw
  viability equivalent to that of standard DMSO-based
  cryopreservation methods, (2) postthaw viability can be
  significantly increased through the use of an
  intracellular-type solution in conjunction with DMSO, (3)
  the use of HTS allows for cryopreservation to be accomplished with reduced
  levels of cryoprotectants, and (4) the regulation of apoptosis is essential
  for the improvement of cryopreservation outcome.

Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=15805