Stromal-Cell-Derived Extracellular Matrix Promotes the Proliferation and Retains the Osteogenic Differentiation Capacity of Mesenchymal Stem Cells on Three-Dimensional Scaffolds

Abstract

To date, expansion of bone-marrow-derived mesenchymal stem cells (MSCs) is typically carried out on twodimensional (2D) tissue culture plastic. Since this 2D substratum is very different from the physiological situation, MSCs gradually lose their unique multipotent properties during expansion. Recently, the role of the extracellular matrix (ECM) microenvironment (''niche'') in facilitating and regulating stem cell behavior in vivo has been elucidated. As a result, investigators have shifted their efforts toward developing threedimensional (3D) scaffolds capable of functioning like the native tissue ECM. In this study, we demonstrated that stromal-cell-derived ECM, formed within a collagen/hydroxyapatite (Col/HA) scaffold to mimic the bone marrow ''niche,'' promoted MSC proliferation and preserved their differentiation capacity. The ECM was synthesized by MSCs to reconstitute the tissue-specific 3D microenvironment in vitro. Following deposition of the ECM inside Col/HA scaffold, the construct was decellularized and reseeded with MSCs to study their behavior. The data showed that MSCs cultured on the ECM-Col/HA scaffolds grew significantly faster than the cells from the same batch cultured on the regular Col/HA scaffolds. In addition, MSCs cultured on the ECMCol/HA scaffolds retained their ''stemness'' and osteogenic differentiation capacity better than MSCs cultured on regular Col/HA scaffolds. When ECM-Col/HA scaffolds were implanted into immunocompromised mice, with or without loading MSCs, it was found that those scaffolds formed less bone as compared with regular Col/ HA scaffolds (i.e., without ECM), in both cases of with or without loading MSCs. The in vivo study further confirmed that the ECM-Col/HA scaffold was a suitable mimic of the bone marrow ''niche.'' This novel 3D stromal-cell-derived ECM system has the potential to be developed into a biomedical platform for regenerative medicine applications.

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

“…Purified ECM proteins (Ng et al, 2016;Santiago et al, 2009) and decellularised matrices derived from tissues (Agmon and Christman, 2016;Bourgine et al, 2013;Swinehart and Badylak, 2016) and cells (Fitzpatrick and McDevitt, 2015) [e.g. permanently differentiated cells (Bourget et al, 2012;Bourget et al, 2017;Kaukonen et al, 2017;Scherzer et al, 2015), mesenchymal stem cells (Antebi et al, 2015;Cai et al, 2015;Shakouri-Motlagh et al, 2017;Thakkar et al, 2013;Zeitouni et al, 2012), immortalised cells (Kusuma et al, 2017) and cell lines (Hoshiba and Gong, 2018)], alone or in combination with a three-dimensional scaffold, are potent regulators of cell fate. Considering that, in traditional culture systems, ECM deposition is very slow and MMC significantly increases and accelerates ECM deposition, the present study ventured to assess the influence of MMC in human BMSCs that were cultured in the presence or absence of MMC and, subsequently, differentiated towards chondrogenic, adipogenic and osteogenic lineage, again, in the presence or absence of MMC.…”

One size does not fit all: developing a cell-specific niche for in vitro study of cell behavior

“…It is very doubtful that the intricate and highly ordered nature of the niche can be reproduced with synthetic or purified components. In an effort to reconstitute the MSC niche in vitro, we previously described a cell-assembled ECM that recapitulates the native microenvironment and dramatically promotes MSC replication and facilitates the retention of their multipotency [12,26,27]. …”

Mesenchymal Stem Cells Reconditioned in Their Own Serum Exhibit Augmented Therapeutic Properties in the Setting of Acute Respiratory Distress Syndrome

“…The key findings of this study are (a) MSC treatment appears to suppress proinflammatory cytokine levels, whereas promoting secretion of anti-inflammatory mediators following administration in vivo; (b) ex vivo experiments reflect trends seen in vivo, in which the IL-6 and IL-1RA relationship appears to play a prominent role; and (c) MSCs reconditioned in their own serum exhibit augmented immunotherapeutic function, which appears to be mediated in part by TLR-4, VEGF, and PGE 2 . Given emerging promise of MSC therapy for ARDS [9,17], we sought to characterize the ARDS microenvironment and evaluate MSC function in a subset of animal models from a larger study, in which ARDS was induced by smoke inhalation and severe burns [16,17]. MSC infusion to human patients has been shown to improve resolution of respiratory and multiorgan failure through reducing pulmonary and systemic markers of inflammation [18].…”