Decellularized ECM effects on human mesenchymal stem cell stemness and differentiation

Abstract

Microenvironment extracellular matrices (ECMs) influence cell adhesion, proliferation and differentiation. The ECMs of different microenvironments have distinctive compositions and architectures. This investigation addresses effects ECMs deposited by a variety of cell types and decellularized with a cold-EDTA protocol have on multipotent human mesenchymal stromal/stem cell (hMSC) behavior and differentiation. The cold-EDTA protocol removes intact cells from ECM, with minimal ECM damage and contamination. The decellularized ECMs deposited by cultured hMSCs, osteogenic hMSCs, and two smooth muscle cell (SMC) lines were tested for distinctive effects on the behavior and differentiation of early passage (‘naïve’) hMSC plated and cultured on the decellularized ECMs. Uninduced hMSC decellularized ECM enhanced naïve hMSC proliferation and cell motility while maintaining stemness. Decellularized ECM deposited by osteogenic hMSCs early in the differentiation process stimulated naïve hMSCs osteogenesis and substrate biomineralization in the absence of added dexamethasone, but this osteogenic induction potential was lower in ECMs decellularized later in the osteogenic hMSC differentiation process. Decellularized ECMs deposited by two smooth muscle cell lines induced naïve hMSCs to become smooth muscle cell-like with distinctive phenotypic characteristics of contractile and synthetic smooth muscle cells. This investigation demonstrates a useful approach for obtaining functional cell-deposited ECM and highlights the importance of ECM specificity in influencing stem cell behavior.

Effects of Human Fibroblast-Derived Extracellular Matrix on Mesenchymal Stem Cells

“…Moreover, they have shown that MSCs cultured on osteogenic hMSC-or osteoblast-derived ECM are directed toward osteoblasts while MSCs on chondrocyte-or smooth muscle cell-derived ECM are induced into chondrocyte or smooth muscle cell, respectively. Rao et al have also shown that the osteoinductive potential of ECM produced by osteogenic hMSCs in early differentiation stage is greater than that in late differentiation stage, indicating that the ECM compositions at distinct differentiation stages are different [38].…”

“…*p < 0.05; **p < 0.01; n = 3. Scale bar: 200 μm [37,38]. Moreover, they have shown that MSCs cultured on osteogenic hMSC-or osteoblast-derived ECM are directed toward osteoblasts while MSCs on chondrocyte-or smooth muscle cell-derived ECM are induced into chondrocyte or smooth muscle cell, respectively.…”

Decellularized extracellular matrix of human umbilical vein endothelial cells promotes endothelial differentiation of stem cells from exfoliated deciduous teeth

“…One possible reason could be lack of environmental signaling cues within the suboptimal two‐dimensional (2‐D) ex vivo culture milieu that compromises cellular responses and their differentiation potentials. Physiologically in vivo , MSCs reside in a three‐dimensional (3‐D) microenvironment surrounded by extracellular matrix (ECM) which is composed of a diverse and complex array of molecules that orchestrate cellular and molecular events during tissue development, repair, and remodeling . While various singular ECM components such as fibronectin, collagen, laminin, or the animal‐derived mixture matrigel have been employed as surface coatings to promote specific cellular biological functions, these commercially available products alone fail to replicate the compositional and structural complexity of natural‐occurring ECM and may provoke adverse host immunological responses due to the animal origins of some of these molecules.…”

“…Physiologically in vivo , MSCs reside in a three‐dimensional (3‐D) microenvironment surrounded by extracellular matrix (ECM) which is composed of a diverse and complex array of molecules that orchestrate cellular and molecular events during tissue development, repair, and remodeling . While various singular ECM components such as fibronectin, collagen, laminin, or the animal‐derived mixture matrigel have been employed as surface coatings to promote specific cellular biological functions, these commercially available products alone fail to replicate the compositional and structural complexity of natural‐occurring ECM and may provoke adverse host immunological responses due to the animal origins of some of these molecules. In contrasts, human‐derived ECM substrata, after removal of antigenic cellular components by decellularization could preserve the inherent ultrastructure and matrix components of native ECM including proteins, proteoglycans and glycosaminoglycans (GAG), thereby creating a more biomimetic microenvironment facilitating stem cell adhesion, migration and proliferation, whilst maintaining their potency in specific cell‐lineage differentiation …”

Growth plate extracellular matrix-derived scaffolds for large bone defect healing

“…Native and in vitro cell culture derived ECMs are commonly used to produce scaffolds for tissue engineering (Almeida et al, 2014;Cunniffe et al, 2015;Jin et al, 2007;Kheir et al, 2011;Kim et al, 2015;Lau et al, 2012;Lu et al, 2011;Pati et al, 2015;Pattabhi et al, 2014;Sutherland et al, 2015); however, the ideal matrix to facilitate bone regeneration has yet to be established. As previously stated, the GP matrix contains many important growth factors for inducing bone growth which render it as an attractive option for generating an osteoinductive ECM derived scaffold.…”