Molecular- and microarray-based analysis of diversity among resting and osteogenically induced porcine mesenchymal stromal cells of several tissue origin

Abstract

Mesenchymal stromal cells (MSCs) play a pivotal role in modern therapeutic approaches in bone‐healing disorders. Although bone marrow‐derived MSCs are most frequently used, the knowledge that many other adult tissues represent promising sources for potent MSCs has gained acceptance. In the present study, the osteogenic differentiation potential of porcine skin fibroblasts (FBs), as well as bone marrow‐ (BMSCs), adipose tissue‐ (ASCs) and dental pulp‐derived stromal cells (DSCs) were evaluated. However, additional application of BMP‐2 significantly elevated the delayed osteogenic differentiation capacity of ASC and FB cultures, and in DSC cultures the supplementation of platelet‐rich plasma increased osteogenic differentiation potential to a comparable level of the good differentiable BMSCs. Furthermore, microarray gene expression performed in an exemplary manner for ASCs and BMSCs revealed that ASCs and BMSCs use different gene expression patterns for osteogenic differentiation under standard media conditions, as diverse MSCs are imprinted dependent from their tissue niche. However, after increasing the differentiation potential of ASCs to a comparable level as shown in BMSCs, a small subset of identical key molecules was used to differentiate in the osteogenic lineage. Until now, the importance of identified genes seems to be underestimated for osteogenic differentiation. Apparently, the regulation of transmembrane protein 229A, interleukin‐33 and the fibroblast growth factor receptor‐2 in the early phase of osteogenic differentiation is needed for optimum results. Based on these results, bone regeneration strategies of MSCs have to be adjusted, and in vivo studies on the osteogenic capacities of the different types of MCSs are warranted. Copyright © 2016 The Authors Tissue Engineering and Regenerative Medicine published by John Wiley & Sons, Ltd.

Osteogenic differentiation of human mesenchymal stromal cells and fibroblasts differs depending on tissue origin and replicative senescence

“…Medium was replaced twice a week. Alternatively, OM was supplemented with 450 ng/ml BMP-2 (PeproTech, Hamburg, Germany) 17 . After 0, 7, 14 and 21 days, the osteogenic differentiation capacity was determined using the alizarin red S assay as described below.…”

“…This phenomenon is underlined by the fact that inconsistent studies describe different effects of BMP-2 and TGF-β, which can induce corresponding pathways. High doses of BMP-2 can accelerate osteogenic differentiation in BMSC or pig ASC 17 , 18 . In stem cells from individuals with Marfan syndrome, exogenous BMP-2 supplementation antagonized TGF-β signaling and rescued the ability to differentiate osteogenically 19 .…”

Cryopreserved H2 O2 -preconditioned human adipose-derived stem cells exhibit fast post-thaw recovery and enhanced bioactivity against oxidative stress

“…In the case of chondrogenic differentiation, cells were seeded at 15,000 cells/cm 2 , and mineralized nodules were qualitatively assessed with Alcian blue (Sigma; Bayraktar et al, ). Samples were fixed in 4% formaldehyde for 15 min at room temperature (RT), washed first with PBS, followed by distilled water and HCl 0.1 N (pH 1), and incubated with Alcian blue 1% in HCl 0.1 N (pH 1) for 2 hr at RT in the darkness.…”

Adipose-Derived Stem Cells in Bone Tissue Engineering: Useful Tools with New Applications

“…PRP can be used to induce the osteogenic differentiation of ASCs, both in vitro and in vivo [132, 154]. Several authors incorporated PRP into different types of scaffold in order to have a controlled and prolonged release of GFs and to favor ASC-mediated bone formation [155–158]. These systems allowed an increased osteogenic capacity of ASCs and an enhanced bone deposition, both in vitro and in animal models.…”