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Cell-secreted extracellular matrix, independent of cell source, promotes the osteogenic differentiation of human stromal vascular fraction.

ABSTRACT: Lipoaspirates contain a readily accessible heterogeneous cell source for use in bone regeneration collectively referred to as the stromal vascular fraction (SVF). However, the osteogenic potential of SVF is inferior to other progenitor cell populations, thereby requiring alternative strategies to potentiate its effective use in cell-based therapies of bone repair. Cell-secreted extracellular matrix (ECM) is a promising substrate to guide cell phenotype or for use in biomaterial design, yet the instructional capacity of ECMs produced by various cell types is unknown. To determine whether the bioactivity of cell-secreted ECM was dependent on cell source, we assessed the osteogenic response of human SVF on ECMs secreted by bone marrow-derived mesenchymal stem cells (MSCs), adipose stromal cells (ASCs), and human dermal fibroblasts (HDFs). Tissue culture plastic (TCP), type I collagen, and ECM induced expression of integrin subunits ?2, ?5, and ?1 in SVF, yet seeding efficiency was only improved on MSC-derived ECM. Regardless of ECM source, SVF deposited over 8- and 1.3-fold more calcium compared to TCP and collagen-coated controls, respectively. Flow cytometry confirmed that SVF cultured on ECM retained CD31 and CD34 positive cell populations better than TCP. After depleting accessory cells, ASCs deposited significantly less calcium compared to donor-matched SVF. This function was partially restored in the presence of MSC-derived ECM when donor-matched endothelial cells (ECs) were added in an ASC/EC co-culture, confirming a role for ECs in osteogenic differentiation. These findings support the use of cell-derived ECM as a means to promote cell retention and osteogenic differentiation of SVF.

SUBMITTER: Harvestine JN

PROVIDER: S-EPMC6261520 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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Cell-secreted extracellular matrix, independent of cell source, promotes the osteogenic differentiation of human stromal vascular fraction.

Harvestine Jenna N JN Orbay Hakan H Chen Jonathan Y JY Sahar David E DE Leach J Kent JK

Journal of materials chemistry. B 20180529 24

Lipoaspirates contain a readily accessible heterogeneous cell source for use in bone regeneration collectively referred to as the stromal vascular fraction (SVF). However, the osteogenic potential of SVF is inferior to other progenitor cell populations, thereby requiring alternative strategies to potentiate its effective use in cell-based therapies of bone repair. Cell-secreted extracellular matrix (ECM) is a promising substrate to guide cell phenotype or for use in biomaterial design, yet the i ...[more]

PMID: 30505446

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Project description:Background: Ectopic vascular calcifications represent a major clinical problem associated with cardiovascular disease and mortality. However, the mechanisms underlying pathological vascular calcifications are largely unknown hampering the development of therapies to tackle this life threatening medical condition. Results: In order to gain insight into the genes and mechanisms driving this pathological calcification process we analyzed the transcriptional profile of calcifying vascular smooth muscle cells (C-VSMCs). These profiles were compared to differentiating osteoblasts, cells that constitute their physiological calcification counterparts in the body. Overall the transcriptional program of C-VSMC and osteoblasts did not overlap. Several genes, some of them relevant for bone formation, were distinctly modulated by C-VSMCs which did not necessarily lose their smooth muscle cell markers while calcifying. Bioinformatics gene clustering and correlation analysis disclosed limited bone-related mechanisms being shared by two cell types. Extracellular matrix (ECM) and biomineralization genes represented common denominators between pathological vascular and physiological bone calcifications. These genes constitute the strongest link between these cells and represent potential drivers for their shared end-point phenotype. Conclusions: the analyses support the hypothesis that VSMC trans-differentiate into C-VSMCs keeping their own identity while using mechanisms that osteoblasts use to mineralize. The data provide novel insights into groups of genes and biological processes shared in MSC and VSMC osteogenic differentiation. The distinct gene regulation between C-VSMC and osteoblasts might hold clues to find cell-specific pathway modulations, opening the possibility to tackle undesired vascular calcifications without disturbing physiologic bone formation and vice versa. Total RNA obtained from hMSC and hVSMC cultured in osteogenic differentiation medium supplemented with 1.8 mM Ca2+ for 0, 2, 8, 12 or 25 days respectively. For each timepoint 3 replicates were used, with exception for day 0 where 4 replicates were collected.

Dataset Information

Cell-secreted extracellular matrix, independent of cell source, promotes the osteogenic differentiation of human stromal vascular fraction.

Publications

Cell-secreted extracellular matrix, independent of cell source, promotes the osteogenic differentiation of human stromal vascular fraction.

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