Cell-derived decellularized extracellular matrices

Abstract

The ability to create cell-derived decellularized matrices in a dish gives researchers the opportunity to possess a bioactive, biocompatible material made up of fibrillar proteins and other factors that recapitulates key features of the native structure and composition of in vivo microenvironments. By using cells in a culture system to provide a natural ECM, decellularization allows for a high degree of customization through the introduction of selected proteins and soluble factors. The culture system, culture medium, cell types, and physical environments can be varied to provide specialized ECMs for wide-ranging applications to study cell-ECM signaling, cell migration, cell differentiation, and tissue engineering purposes. This chapter describes a procedure for performing a detergent and high pH-based extraction that leaves the native, cell-assembled ECM intact while removing cellular materials. We address common evaluation methods for assessing the ECM and its composition as well as potential uses for a decellularized ECM.

Micropatterning Decellularized ECM as a Bioactive Surface to Guide Cell Alignment, Proliferation, and Migration

“…Substrates were decellularized as previously described [ 11 , 13 ]. In brief, the substrates were washed twice with 1× PBS and rinsed twice with wash buffer solution containing 0.1 M sodium phosphate (Na 2 HPO 4 ), 2 mM magnesium chloride (MgCl 2 ), and 2 mM Egtazic acid (EGTA), adjusted to a pH of 9.6.…”

“…Biomimetic structures, in particular, strive to recreate naturally regenerative microenvironments to promote desired cell behavior and mitigate inflammatory responses [ 9 ]. Recently, the extracellular matrix (ECM) has been targeted as a bioactive substrate for tissue repair [ 10 , 11 , 12 , 13 ]. A naturally cell-assembled ECM is able to provide physical support to tissue while concurrently mediating cell and tissue behavior [ 14 , 15 ].…”

“…To create the ECM, the secretion and organization of the macromolecules are mediated by cells themselves [ 18 ]. For FN in particular, as dimers are secreted, they are assembled into a 3D ECM by cellular tension and capable of binding other components [ 11 , 13 , 15 , 18 ]. The cell-secreted ECM then has the potential to be decellularized to remove cells, leaving only a cell-assembled decellularized ECM (dECM).…”

Carrageenan enhances chondrogenesis and osteogenesis in human bone marrow stem cell culture

“…Further, it is generally recognised that in vitro induction is necessary prior to implantation to ensure uniform differentiation and avoid unwanted differentiation, uncontrolled proliferation and senescence Heng et al, 2004;Herberts et al, 2011;Lepperdinger et al, 2008;Trombi et al, 2016;Yorukoglu et al, 2017;Young et al, 2013). Decellularised stem, permanently differentiated and immortalised cell-derived matrices are traditionally used to either support efficient stem cell expansion or to direct stem cells towards a specific lineage (Cheng et al, 2014;Fitzpatrick and McDevitt, 2015;Harris et al, 2018;Kaukonen et al, 2017;Kusuma et al, 2017). However, again due to slow and low level of ECM deposition in traditional in vitro systems, among others, cell-ECM interactions and interconnectivity are not mimicked, leading to less and often uncontrollable differentiation/unwanted trans-differentiation (Choi et al, 2012;Huang et al, 2017;Kwon et al, 2013;Marinkovic et al, 2016).…”

Selection of the Optimal Protocol for Preparation of a Decellularized Extracellular Matrix of Human Adipose Tissue-Derived Mesenchymal Stromal Cells

“…2Е, Ж). Подобные результаты были получены в работе Харриса с коллегами на эмбриональных фибробластах мыши [16]. Молекулы коллагена в составе дцВКМ не выявлялись, что соотносится с данными, полученными на клеточных препаратах, когда иммуноцитохимически был обнаружен коллаген типа I, локализованный внутриклеточно в форме проколлагена.…”