Decellularization of rat adipose tissue, diaphragm, and heart: a comparison of two decellularization methods
Decellularization is a process that involves the removal of cellular material from the tissues and organs while maintaining the structural, functional, and mechanical properties of extracellular matrix. The purpose of this study was to carry out decellularization of rat adipose tissue, diaphragm, and heart by using two different methods in order to compare their efficiency and investigate proliferation profiles of rat adipose-tissue-derived mesenchymal stem cells (AdMSCs) on these scaffolds. Tissues were treated with an optimized detergent-based decellularization (Method A) and a freeze-and-thaw-based decellularization (Method B). AdMSCs were then seeded on scaffolds having a density of 2 × 105 cells/scaffold and AO/PI double-staining and MTT assays were performed in order to determine cell viability. In this study, which is the first research comparing two methods of decellularization of an adipose tissue, diaphragm, and heart scaffolds with AdMSCs, Method A provided efficient decellularization in these three tissues and it was shown that these porous scaffolds were cyto-compatible for the cells. Method B caused severe tissue damage in diaphragm and insufficient decellularization in heart whereas it also resulted in cyto-compatible adipose tissue scaffolds.
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- Akbay E, Onur MA (2018). Myocardial tissue engineering: a
comparative study of different solutions for use as a natural
scaffold being of heart. J Biomed Mater Res (in press).
- Brown BN, Freund JM, Han L, Rubin JP, Reing JE, Jeffries EM, Wolf
MT, Tottey S, Barnes CA, Ratner BD et al. (2011). Comparison
of three methods for the derivation of a biologic scaffold
composed of adipose tissue extracellular matrix. Tissue Eng
Part C-Me 17: 411-421.
- Chang Y, Tsai CC, Liang HC, Sung HW (2002). In vivo evaluation of
cellular and acellular bovine pericardia fixed with a naturally
occurring crosslinking agent (genipin). Biomaterials 23: 2447-
2457.
- Chen S, Li J, Dong P (2013). Utilization of pulsatile flow to
decellularize the human umbilical arteries to make small-
caliber blood vessel scaffolds. Acta Cardiol Sin 29: 451.
- Cheng CW, Solorio LD, Alsberg E (2014). Decellularized tissue and
cell-derived extracellular matrices as scaffolds for orthopaedic
tissue engineering. Biotechnol Adv 32: 462-484.
- Crapo PM, Gilbert, TW, Badylak, SF (2011). An overview of tissue
and whole organ decellularization processes. Biomaterials 32:
3233-3243.
- Dai R, Wang, Z, Samanipour R, Koo KI, Kim K (2016). Adipose-
derived stem cells for tissue engineering and regenerative
medicine applications. Stem Cells Int 2016: 6737345.
- Dunne LW, Huang Z, Meng W, Fan X, Zhang N, Zhang Q, An, Z
(2014). Human decellularized adipose tissue scaffold as a
model for breast cancer cell growth and drug treatments.
Biomaterials 35: 4940-4949.
- Ferrando PM, Balmativola D, Cambieri I, Scalzo MS, Bergallo M,
Annaratone L, Casarin S, Fumagalli M, Stella M, Sapino A et al.
(2016). Glycerolized reticular dermis as a new human acellular
dermal matrix: an exploratory study. PLoS one 11: e0149124.
- Flynn L (2010). The use of decellularized adipose tissue to provide an
inductive microenvironment for the adipogenic differentiation
of human adipose-derived stem cells. Biomaterials 31: 4715-
4724.
- Gilpin A, Yang Y (2017). Decellularization strategies for regenerative
medicine: from processing techniques to applications. Biomed
Res Int 2017: 9831534.
- Gubareva EA, Sjöqvist S, Gilevich IV, Sotnichenko AS, Kuevda EV,
Lim ML, Feliu N, Lemon G, Danilenko KA, Nakokhov RZ et
al. (2016). Orthotopic transplantation of a tissue engineered
diaphragm in rats. Biomaterials 77: 320-335.
- Hoshiba T, Chen G, Endo C, Maruyama H, Wakui M, Nemoto E,
Kawazoe N, Tanaka M (2015). Decellularized extracellular
matrix as an in vitro model to study the comprehensive roles
of the ECM in stem cell differentiation. Stem Cells Int 2016:
6397820.
- Hoshiba T, Lu H, Kawazoe N, Chen G (2010). Decellularized matrices
for tissue engineering. Expert Opin Biol Ther 10: 1717-1728.
- Kajbafzadeh AM, Tafti SHA, Khorramirouz R, Sabetkish S, Kameli
SM, Orangian S, Rabbani S, Oveisi N, Golmohammadi
M, Kashani Z (2017). Evaluating the role of autologous
mesenchymal stem cell seeded on decellularized pericardium
in the treatment of myocardial infarction: an animal study. Cell
Tissue Bank 18: 527-538.
- Mazza G, Rombouts K, Rennie Hall A, Urbani L, Vinh Luong T, Al-
Akkad W, Longato L, Brown D, Maghsoudlou P, Dhillon AP
(2015). Decellularized human liver as a natural 3D-scaffold for
liver bioengineering and transplantation. Sci Rep 5: 13079.
- Niyaz M, Gürpınar ÖA, Günaydın S, Onur MA (2012). Isolation,
culturing and characterization of rat adipose tissuederived
mesenchymal stem cells: a simple technique. Turk J Biol 36.
- Palpant NJ, Metzger JM (2010). Aesthetic cardiology: adipose-
derived stem cells for myocardial repair. Curr Stem Cell Res
Ther 5: 145-152.
- Piccoli M, Urbani L, Alvarez-Fallas ME, Franzin C, Dedja A,
Bertin E, Zuccolotto G, Rosato A, Pavan P, Elvassore N et al.
(2016). Improvement of diaphragmatic performance through
orthotopic application of decellularized extracellular matrix
patch. Biomaterials 74: 245-255.
- Price AP, Godin LM, Domek A, Cotter T, D’Cunha J, Taylor DA,
Panoskaltsis-Mortari A (2014). Automated decellularization of
intact, human-sized lungs for tissue engineering. Tissue Eng
Part C-Me 21: 94-103.
- Rana D, Zreiqat H, Benkirane‐Jessel N, Ramakrishna S, Ramalingam,
M (2017). Development of decellularized scaffolds for stem
cell‐driven tissue engineering. J Tissue Eng Reg Med 11: 942-
965.
- Rigogliuso S, Pavia FC, Brucato V, Carrubba V, Favia P, Intranuovo
F, Gristina R, Ghersi G (2012). Use of modified 3D scaffolds
to improve cell adhesion and drive desired cell responses.
Chemical Engineering Transactions 27: 415-420.
- Sellaro TL, Ravindra AK, Stolz DB, Badylak SF (2007). Maintenance
of hepatic sinusoidal endothelial cell phenotype in vitro using
organ-specific extracellular matrix scaffolds. Tissue Eng 13:
2301-2310.
- Sierad LN, Shaw EL, Bina A, Brazile B, Rierson N, Patnaik SS,
Kennamer A, Odum R, Cotoi O, Terezia P et al. (2015).
Functional heart valve scaffolds obtained by complete
decellularization of porcine aortic roots in a novel differential
pressure gradient perfusion system. Tissue Eng Part C-Me 21:
1284-1296.
- Tapias LF, Ott HC (2014). Decellularized scaffolds as a platform for
bioengineered organs. Curr Opin Organ Transplant 19: 145.
- Thevenot P, Nair A, Dey J, Yang J, Tang L (2008). Method to
analyze three-dimensional cell distribution and infiltration in
degradable scaffolds. Tissue Eng Part C-Me 14:319-331.
- Tığlı RS, Ghosh S, Laha MM, Shevde NK, Daheron L,Gimble J,
Gümüşderelioğlu M and Kaplan DL (2009). Comparative
chondrogenesis of human cell sourcesin 3D scaffolds. J Tissue
Eng Regen Med 3: 348-360.
- Wang L, Johnson JA, Chang DW, Zhang Q (2013a). Decellularized
musculofascial extracellular matrix for tissue engineering.
Biomaterials 34: 2641-2654.
- Wang L, Johnson JA, Zhang Q, Beahm EK (2013b). Combining
decellularized human adipose tissue extracellular matrix and
adipose-derived stem cells for adipose tissue engineering. Acta
Biomaterialia 9: 8921-8931.
- Wei HJ, Chen SC, Chang Y, Hwang SM, Lin WW, Lai PH, Chiang
HK, Hsu LF, Yang HH, Sung HW (2006). Porous acellular
bovine pericardia seeded with mesenchymal stem cells as a
patch to repair a myocardial defect in a syngeneic rat model.
Biomaterials 27: 5409-5419.