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Methods Mol Biol. 2017;1612:15-27. doi: 10.1007/978-1-4939-7021-6_2.

Preparation of Decellularized Biological Scaffolds for 3D Cell Culture.

Author information

1
Department of Bioengineering, University of Pittsburgh, Bridgeside Point II Building, 450 Technology Drive, Suite 300, Pittsburgh, PA, 15219, USA. brownb@upmc.edu.
2
McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA. brownb@upmc.edu.
3
Department of Obstetrics, Gynecology and Reproductive Sciences, Magee-Womens Research Institute, University of Pittsburgh, Pittsburgh, PA, USA. brownb@upmc.edu.
4
Department of Bioengineering, University of Pittsburgh, Bridgeside Point II Building, 450 Technology Drive, Suite 300, Pittsburgh, PA, 15219, USA.

Abstract

The biggest challenge of designing and implementing an in vitro study is developing a microenvironment that most closely represents the interactions observed in vivo. Decellularization of tissues and organs has been shown to be an effective method for the removal of potentially immunogenic constituents while preserving essential growth factors and extracellular matrix (ECM ) proteins necessary for proper cell function. Enzymatic digestion of decellularized tissues allows these tissue-specific components to be reconstituted into bioactive hydrogels through a physical crosslinking of collagen. In the following protocol, we describe unique decellularization methods for both dermis and urinary bladder matrix (UBM) derived from porcine tissues. We then provide details for hydrogel formation and subsequent three-dimensional (3D) culture of two cell types: NIH 3T3 fibroblasts and C2C12 myoblasts .

KEYWORDS:

Cell culture; Decellularization; Extracellular matrix; Hydrogel; Scaffolds

PMID:
28634932
DOI:
10.1007/978-1-4939-7021-6_2
[Indexed for MEDLINE]

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