samples should be stored in a dry environment to avoid moisture uptake during storage, which can result in a decrease of the
glass transition temperature causing degradation reactions in
the sample.
9. Buffer can be replaced several times with fresh buffer to remove
the protectants. Sucrose, however, is a relatively harmless protectant that does not need to be completely removed prior to
transplantation.
10. Alternatively, samples can be manually transferred in the different solutions.
11. Hematoxylin has a deep blue-purple color and stains proteins
nonspecifically. In a typical tissue, nuclei are stained blue,
whereas cytoplasm and extracellular matrix have varying
degrees of pink staining.
References
1. Hammermeister K, Sethi GK, Henderson WG,
Grover FL, Oprian C, Rahimtoola SH (2000)
Outcomes 15 years after valve replacement
with a mechanical versus a bioprosthetic valve:
final report of the veterans affairs randomized
trial. J Am Coll Cardiol 36:1152–1158
2. Jayakrishnan A, Jameela SR (1996) Glutaraldehyde as a fixative in bioprostheses and drug
delivery matrices. Biomaterials 17:471–484
3. Cebotari S, Lichtenberg A, Tudorache I,
Hilfiker A, Mertsching H, Leyh R,
Breymann T, Kallenbach K, Maniuc L,
Batrinac A, Repin O, Maliga O, Ciubotaru A,
Haverich A (2006) Clinical application of tissue engineered human heart valves using autologous progenitor cells. Circulation 114:
I132–I137
4. Cebotari S, Tudorache I, Ciubotaru A,
Boethig D, Sarikouch S, Goerler A,
Lichtenberg A, Cheptanaru E, Barnaciuc S,
Cazacu A, Maliga O, Repin O, Maniuc L,
Breymann T, Haverich A (2011) Use of fresh
decellularized allografts for pulmonary valve
replacement may reduce the reoperation rate
in children and young adults: early report. Circulation 124:S115–S123
5. Moore CH, Martelli V, Al-Janabi N, Ross DN
(1975) Analysis of homograft valve failure in
311 patients followed up to 10 years. Ann
Thorac Surg 20:274–281
6. Davies H, Missen GA, Blandford G, Roberts
CI, Lessof MH, Ross DN (1968) Homograft
replacement of the aortic valve: a clinical and
pathologic study. Am J Cardiol 22:195–217
7. Curtil A, Pegg DE, Wilson A (1997) Freeze
drying of cardiac valves in preparation for cellular repopulation. Cryobiology 34:13–22
8. Wang S, Goecke T, Meixner C, Haverich A,
Hilfiker A, Wolkers WF (2012) Freeze-dried
heart valve scaffolds. Tissue Eng 18:517–525
9. Wang S, Oldenhof H, Dai X, Haverich A,
Hilfiker A, Harder M, Wolkers WF (2015)
Sucrose diffusion in decellularized heart valves
for freeze-drying. Tissue Eng 21:922–931
10. Va ´squez-Rivera A, Oldenhof H, Dipresa D,
Goecke T, Kouvaka A, Will F, Haverich A,
Korossis S, Hilfiker A, Wolkers WF (2018)
Use of sucrose to diminish pore formation in
freeze-dried heart valves. Sci Rep 8:12982
11. Goecke T, Theodoridis K, Tudorache I,
Ciubotaru A, Cebotaria S, Ramm R,
Ho ¨ffler K, Sarikouch S, Va ´squez-Rivera A,
Haverich A, Wolkers WF, Hilfiker A (2018)
In vivo performance of freeze-dried decellularized pulmonary heart valve allo- and xenografts
orthotopically implanted into juvenile sheep.
Acta Biomater 68:41–52
Freeze-Dried Heart Valves
739
glass transition temperature causing degradation reactions in
the sample.
9. Buffer can be replaced several times with fresh buffer to remove
the protectants. Sucrose, however, is a relatively harmless protectant that does not need to be completely removed prior to
transplantation.
10. Alternatively, samples can be manually transferred in the different solutions.
11. Hematoxylin has a deep blue-purple color and stains proteins
nonspecifically. In a typical tissue, nuclei are stained blue,
whereas cytoplasm and extracellular matrix have varying
degrees of pink staining.
References
1. Hammermeister K, Sethi GK, Henderson WG,
Grover FL, Oprian C, Rahimtoola SH (2000)
Outcomes 15 years after valve replacement
with a mechanical versus a bioprosthetic valve:
final report of the veterans affairs randomized
trial. J Am Coll Cardiol 36:1152–1158
2. Jayakrishnan A, Jameela SR (1996) Glutaraldehyde as a fixative in bioprostheses and drug
delivery matrices. Biomaterials 17:471–484
3. Cebotari S, Lichtenberg A, Tudorache I,
Hilfiker A, Mertsching H, Leyh R,
Breymann T, Kallenbach K, Maniuc L,
Batrinac A, Repin O, Maliga O, Ciubotaru A,
Haverich A (2006) Clinical application of tissue engineered human heart valves using autologous progenitor cells. Circulation 114:
I132–I137
4. Cebotari S, Tudorache I, Ciubotaru A,
Boethig D, Sarikouch S, Goerler A,
Lichtenberg A, Cheptanaru E, Barnaciuc S,
Cazacu A, Maliga O, Repin O, Maniuc L,
Breymann T, Haverich A (2011) Use of fresh
decellularized allografts for pulmonary valve
replacement may reduce the reoperation rate
in children and young adults: early report. Circulation 124:S115–S123
5. Moore CH, Martelli V, Al-Janabi N, Ross DN
(1975) Analysis of homograft valve failure in
311 patients followed up to 10 years. Ann
Thorac Surg 20:274–281
6. Davies H, Missen GA, Blandford G, Roberts
CI, Lessof MH, Ross DN (1968) Homograft
replacement of the aortic valve: a clinical and
pathologic study. Am J Cardiol 22:195–217
7. Curtil A, Pegg DE, Wilson A (1997) Freeze
drying of cardiac valves in preparation for cellular repopulation. Cryobiology 34:13–22
8. Wang S, Goecke T, Meixner C, Haverich A,
Hilfiker A, Wolkers WF (2012) Freeze-dried
heart valve scaffolds. Tissue Eng 18:517–525
9. Wang S, Oldenhof H, Dai X, Haverich A,
Hilfiker A, Harder M, Wolkers WF (2015)
Sucrose diffusion in decellularized heart valves
for freeze-drying. Tissue Eng 21:922–931
10. Va ´squez-Rivera A, Oldenhof H, Dipresa D,
Goecke T, Kouvaka A, Will F, Haverich A,
Korossis S, Hilfiker A, Wolkers WF (2018)
Use of sucrose to diminish pore formation in
freeze-dried heart valves. Sci Rep 8:12982
11. Goecke T, Theodoridis K, Tudorache I,
Ciubotaru A, Cebotaria S, Ramm R,
Ho ¨ffler K, Sarikouch S, Va ´squez-Rivera A,
Haverich A, Wolkers WF, Hilfiker A (2018)
In vivo performance of freeze-dried decellularized pulmonary heart valve allo- and xenografts
orthotopically implanted into juvenile sheep.
Acta Biomater 68:41–52
Freeze-Dried Heart Valves
739