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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