Table 1.
Comparison of decellularization methods investigated.
| Decellularization Solution | Nuclease Solutions | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Treatment Group |
Number of Sonications |
Time in Decell Solution |
70 % EtOH Treatment Time |
EDTA | Triton X-100 | Deoxycholic Acid | Sodium Azide | SDS | DNase | RNase | Summary of Findings |
| Decell 1 | 6 | 3 | - | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 7.19 ng/mg Average [GAG): 52.63 μg/mg Histology: Visable Nuclei and Faded Blue ECM |
| Decell 2 | 12 | 3 | - | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 15.26 ng/mg Average [GAG]: 59.81 μg/mg |
| Decell 3 | 6 | 6 | - | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 10.21 ng/mg Average [GAG]: 70.67 μg/mg |
| Decell 4 | 12 | 6 | - | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 7.53ng/mg Average [GAG]: 63.04 μg/mg Histology:Visable Nuclei and Deep Blue ECM |
| Decell 5 | 6 | 3 | - | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 1440 mU/mL | 1440 mU/mL | Average [DNA]: 11.59 ng/mg Average [GAG]: 83.09 μg/mg |
| Decell 6 | 3 | 3 | - | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 10.45 ng/mg Average [GAG]: 57.99 μg/mg |
| Decell 7* | 3 | 3 | - | 0.4% (w/v) | 1.2% (v/v) | 2% (w/v) | 0.04% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 33.61 ng/mg Average [GAG]: 216.98 μg/mg Histology: Empty Lacunae and Faded Blue ECM |
| Decell 8 | 3 | 3 | 30 min | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 15.59 ng/mg Average [GAG]: 108.42 μg/mg |
| Decell 9 | 3 | 3 | 60 min | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 9.07 ng/mg Average [GAG]: 184.53 μg/mg |
| Decell 10* | 3 | 3 | 45 min | 0.2% (w/v) | 0.6% (v/v) | 1% (w/v) | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 33.08 ng/mg Average [GAG]: 208.93 μg/mg Histology: Nuclei Present and Faded Blue ECM |
| Decell 11 | 3 | 3 | - | 0.2% (w/v) | 0.6% (v/v) | - | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 15.22 ng/mg Average [GAG]: 133.59 μg/mg Histology: Nuclei Present and Intense Blue ECM |
| Decell 12 | 3 | 3 | - | 2.21 mM | - | - | - | 0.4% (w/v) | 720 mU/mL | 720 mU/mL | Average [DNA]: 23.31 ng/mg Average [GAG]: 71.92 μg/mg Histology: Nuclei Present and Faded Blue ECM |
| Decell 13* | 3 | 3 | - | 0,4% (w/v) | 1.2% (v/v) | - | 0.02% (w/v) | - | 720 mU/mL | 720 mU/mL | Average [DNA]: 9.07 ng/mg Average [GAG]: 205.5 μg/mg Histology: Empty Lacunae and Deep Blue ECM |