Skip to main content

Table 1 Sample groups and decellularization protocols

From: Automated freeze-thaw cycles for decellularization of tendon tissue - a pilot study

Group Protocol Decellularization procedures
   5 repetitions of freeze-thawing Further treatment
   Cooling Freeze hold Heating Thaw hold  
1 Auto-Protocol 1 -50 °C per min 3 min at -80 °C +50 °C per min 10 min at +20 °C 48 h distilled water
48 h 1% Triton X-100
Washing steps
2 Auto-Protocol 2 -20 °C per min 3 min at -80 °C +20 °C per min 10 min at +20 °C
3 Manual-Protocol Manual transfer 2 min in liquid nitrogen Manual transfer 10 min in 37 °C PBS
  Control No treatment
  1. Equine superficial digital flexor tendon samples of group 1 and group 2 were processed by automated freeze-thaw cycles, differing in the performed cooling and heating rates (Auto-Protocol 1 and Auto-Protocol 2). Both of the applied cooling and heating rates describe a temperature change per unit time. For Auto-Protocol 1 as well as for Auto-Protocol 2 the maximum reached temperature was + 20 °C (thaw hold for 10 min) and the minimum reached temperature was -80 °C (freeze hold for 3 min). All temperature regulations of the automated freeze-thaw cycles were carried out by a controlled rate freezer (PLANER® Kryo 360–1.7) that utilizes liquid nitrogen to adjust temperature. Group 3 included manual freeze-thaw cycles. Further steps of decellularization were the same for all sample groups. Tendon samples classified as internal control underwent no decellularization