The Rapidly Evolving Concept of Whole Heart Engineering

Table 1

Schematic description of the methodologies applied so far for whole heart decellularization.

Author

Year

Species

Decellularization protocol

Ott

2008

Rat

10 μM adenosine in heparanized PBS, 15 min, 77.4 mmHg

1% SDS in deionized water, 12 h, 77.4 mmHg

Deionized water, 15 min, 77.4 mmHg

1% Triton X-100 in deionized water, 30 min, 77.4 mmHg

100 IU/ml penicillin-G/streptomycin and 100 IU/ml amphotericin B in PBS, 124 h

Wainwright

2010

Pig

Freezing, 16 h, −80°C

Thawing in distilled water, RT

Distilled water, 15 min, 1 l/min, RT

2x PBS, 15 min, 1 l/min, RT

0.02% trypsin, 0.05% EDTA, and 0.05% NaN3, 2 h, 1 l/min, 37°C

Distilled water, 5 min, RT

2x PBS, 15 min, 1 l/min, RT

3% Triton X-100, 0.05% EDTA, and 0.05% NaN3 in distilled water, 2 h, 1.3 l/min, RT

Distilled water, 5 min, RT

2x PBS, 15 min, 1 l/min, RT

4% DCA in distilled water, 2 h, 1.3 l/min, RT

Distilled water, 5 min, RT

2x PBS, 15 min, 1 l/min, RT

0.1% peracetic acid and 4% ethanol, 1 h, 1.7 l/min

PBS, 15+ 15 min, 1.7 l/min

Distilled water, 15+ 15+ 15 min, 1.7 l/min

Weymann

2011

Pig

4% SDS in PBS, 3 h, 2 l/min, 100 mmHg, 37°C

PBS, 15 min, 37°C

4% SDS in PBS, 3 h, 2 l/min, 100 mmHg, 37°C

PBS, 15 min, 37°C

4% SDS in PBS, 3 h, 2 l/min, 100 mmHg, 37°C

PBS, 15 min, 37°C

4% SDS in PBS, 3 h, 2 l/min, 100 mmHg, 37°C

PBS, 24 h, 1.5 l/min

2014

100 μg/ml penicillin/streptomycin in PBS, 24 h, 1.5 l/min

Akhyari

2011

Rat

I

10 mM adenosine in heparanized PBS, 15 min, 77.5 mmHg

1% SDS in deionized water, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

1% Triton X-100 in deionized water, 30 min, 77.5 mmHg

100 IU/ml penicillin-G/streptomycin and amphotericin B, 124 h

II

0.025% trypsin and 0.05% EDTA in PBS, 1 h, 77.5 mmHg, 37°C

Deionized water, 15 min

3% Triton X-100 in deionized water, 1 h, 77.5 mmHg

Deionized water, 15 min

4% DCA in deionized water, 1 h, 77.5 mmHg, RT

Deionized water, 15 min, RT

0.1% acetic acid in deionized water, 1 h

III

20% glycerol, 0.05% NaN3, and 25 mM EDTA in 0.9% NaCl, 3 d, 77.5 mmHg

4.2% DCA and 0.05% NaN3 in deionized water, 3 d, 77.5 mmHg

20% glycerol, 0.05% NaN3, and 25 mM EDTA in 0.9% NaCl, 2 d, 77.5 mmHg

1% SDS and 0.05% NaN3 in deionized water, 2 d, 77.5 mmHg

3% Triton X-100 and 0.05% NaN3 in deionized water, 2 d, 77.5 mmHg

100 IU/ml penicillin-G/streptomycin, 12 h

IV

10 mM adenosine in heparanized PBS, 15 min, 77.5 mmHg

1% SDS, 1% DCA, and 0.05% NaN3 in deionized water, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

20% glycerol, 0.05% NaN3, and 25 mM EDTA in 0.9% NaCl, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

1% Saponin and 0.05% NaN3 in deionized water, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

20% glycerol, 0.05% NaN3, and 25 mM EDTA in 0.9% NaCl, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

200 IU/ml Dnase I and 50 ml/mol MgCl in PBS

Aubin

2013

Rat

10 mM adenosine in heparanized PBS, 15 min, 77.5 mmHg

1% SDS, 1% DCA, and 0.05% NaN3 in deionized water, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

20% glycerol, 0.05% NaN3, and 25 mM EDTA in 0.9% NaCl, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

1% Saponin and 0.05% NaN3 in deionized water, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

20% glycerol, 0.05% NaN3, and 25 mM EDTA in 0.9% NaCl, 12 h, 77.5 mmHg

Deionized water, 15 min, 77.5 mmHg

200 IU/ml Dnase I and 50 mM MgCl in PBS

100 IU/ml penicillin/streptomycin in PBS, 12 h, 4°C

Remlinger

2012

Pig

Freezing, 24 h, −80°C

Thawing in deionized water, ON, 4°C

Deionized water, 15–25 min, 400 ml/min

2x PBS, 15 min, 700 ml/min

Deionized water, 10 min, 750 ml/min

0.2% trypsin, 0.05% EDTA, and 0.05% NaN3, 3 h, 1200–1800 ml/min, 37°C

Deionized water, 10 min, 1900 ml/min, RT

2x PBS, 10 min, 1950 ml/min, RT

3% Triton X-100, 0.05% EDTA, and 0.05% NaN3, 2.5 h, 2000–2100 ml/min, RT

Deionized water, 10 min, 2150 ml/min, RT

2x PBS, 10 min, 2180 ml/min, RT

4% SDC, 3 h, 2200 ml/min, RT

Deionized water, 15 min, 2200 ml/min, RT

2x PBS, 15 min, 2200 ml/min, RT

Deionized water, 5 min, 750 ml/min, RT

1x PBS, 5 min, 1500 ml/min, RT

Merna

2013

Pig

I

Freezing, 24 h, −80°C

Thawing, RT

0.02% Trypsin, 0.05% EDTA, and 0.05% NaN3, 3 d

3% Triton X-100, 0.05% EDTA, and 0.05% NaN3, 4 d

II

Freezing, 24 h, −80°C

Thawing, RT

0.02% Trypsin, 0.05% EDTA, and 0.05% NaN3, 1 d

3% Triton X-100, 0.05% EDTA, and 0.05% NaN3, 6 d

III

Freezing, 24 h, −80°C

Thawing, RT

0.02% Trypsin, 0.05% EDTA, and 0.05% NaN3, 7 d

IV

Freezing, 24 h, −80°C

Thawing, RT

3% Triton X-100, 0.05% EDTA, and 0.05% NaN3, 7 d

Methe

2014

Pig

Washed in 1% penicillin/streptomycin and 1% amphotericin B

Freezing in PBS, 24 h, −80°C

Thawing, RT

Washed in distilled water

Short perfusion of 4% SDC

Immersion and agitation in 4% SDC, 6 h, 37°C

Perfusion with 1% SDC, 6 h, RT

Perfusion with distilled water, 6 h, RT

Perfusion with 1% Triton X-100, 12 h

Immersion and agitatio in 1% Triton X-100, 12 h

Perfusion with 0.1% peracetic acid in PBS, 3 h

Momtahan

2015

Pig

Washed in heparin (10 IU/m), 100 IU/ml penicillin, 100 μg/ml streptomycin, and 25 μg/ml amphotericin B

Freezing in heparanized water, −20°C

Thawing, ON, 4°C

1x PBS, 1 h, 0–4 psi, 23°C

Distilled water, 1 h, 5 psi, 23°C

0.5% SDS in distilled water, 2 h, 5 psi, 23°C

Distilled water, 2 h, 5 psi, 23°C

Distilled water, ON, 3 psi, 23°C

1% Triton X-100, 2 h, 3–5 psi, 23°C

Distilled water, 5 h, 5 psi, 23°C

Guyette

2016

Human

Heparanized PBS (1 IU/ml), 1 h, 60 mmHg, RT

1% SDS in deionized water, 168 h, 60 mmHg, RT

Deionized water, 24 h, 60 mmHg, RT

1% Triton X-100 in deionized water, 24 h, 60 mmHg, RT

PBS, 168 h, 60 mmHg, RT

Kitahara

2016

Pig

Freezing, 24 h, −80°C

Thawing, 4°C

Deionized water, 15 min, 100 ml/min

1% SDS in deionized water, 3 h, 100 mmHg, 200–1000 ml/min, 37°C

Deionized water, 15 min, 1000 ml/min

1% Triton X-100 in deionized water, 3 h, 1000 ml/min

Deionized water, 1 h, 1000 ml/min

Sanchez

2016

Human

1% SDS in deionized water, 60 l in 4 d, 80–100 mmHg

Washed in water, 20 l, 80–100 mmHg

Penicillin/streptomycin in PBS, 10 l, 80–100 mm Hg

RT: room temperature; EDTA: ethylenediaminetetraacetic acid; NaN₃: sodium azide; DCA: deoxycholic acid; PBS: phosphate-buffered saline; SDS: sodium dodecyl sulfate; SDC: sodium deoxycholate; psi: pound-force per square inch. This group of washes has been repeated 8 times. This group of washes has been repeated 3 times. This group of washes has been repeated 3 times.