Characterization of In Vitro Expanded Bone Marrow-Derived Mesenchymal Stem Cells Isolated from Experimental Autoimmune Encephalomyelitis Mice

Abstract

Extensive experimental studies indicate that autologous bone marrow mesenchymal stem cells (BMSCs) are able to ameliorate experimental autoimmune encephalomyelitis (EAE) and potentially multiple sclerosis. However, the impact that the inflammatory environment present in EAE may have on the biological properties of BMSCs expanded in vitro for transplantation is yet to be clarified. It was investigated whether BMSCs isolated from EAE-induced C57bl6/J mice and expanded in vitro preserve the properties of BMSCs isolated from healthy donors (BMSCs-control). The mesenchymal origin, the differentiation potential, and the transcriptional expression profile of six histone-modifying genes were studied in both groups of BMSCs. BMSCs-EAE exhibited distinct morphology and larger size compared to BMSCs-control, higher degree of proliferation and apoptosis, differences in the adipogenesis and the osteogenesis induction, and differential expression of stromal markers and markers of progenitor and mature neuronal/glial cells. Moreover, BMSCs-EAE exhibited different expression patterns on a number of histone-modifying genes compared to controls. We recorded manifold differences, both phenotypical and functional, of in vitro expanded BMSCs-EAE in comparison to their healthy donor-derived counterparts that may be attributed to the inflammatory environment they originated from. Whether our findings may be of any clinical relevance needs to be clarified in future studies, in vivo.

CNS disease diminishes the therapeutic functionality of bone marrow mesenchymal stem cells

“…This study isolated MSCs from EAE mice early on in the disease, when the mice showed clinical symptoms such as tail paralysis or hind limb weakness. In contrast, a more recent study compared BM-MSCs isolated from MOG EAE mice later in disease, when symptoms are more severe, and found that these EAE-MSCs were different from naïve MSCs in terms of growth rate, differentiation potential, and mRNA expression levels of important histone-modifying genes (Zacharaki et al, 2013). This later study did not compare EAE-MSCs to naïve MSCs in terms of therapeutic efficacy.…”

“…To assess proliferation, MSC treated with 10 uM BrdU (Sigma) for sixteen hours. Coverslips were then fixed for 10 minutes with 4% PFA and stained for rat anti-BrdU (Abcam: ab6326) using a previously described protocol (Zacharaki et al, 2013). …”

Bone Mesenchymal Stem Cells Contributed to the Neointimal Formation after Arterial Injury

“…A panel of genetic, antigenic and functional assays is required to provide optimal characterization of BMSCs[38]. At first, we demonstrated that BMSCs in vitro had EC characteristics phenotypes and angiogenic properties based on the following observations.…”

Mesenchymal Stem Cells and Induced Pluripotent Stem Cells as Therapies for Multiple Sclerosis

“…BM-MSCs isolated from EAE mice exhibited distinct morphology, elevated ratio of proliferation and apoptosis, differences in the adipogenesis and the osteogenesis induction, distinct expression profile of stromal markers [26] and different expression patterns on six histone-modifying genes compared to MSCs from control mice [27]. However, another report indicated that the inflammatory process did not exert any deleterious effect on the functional/biological properties of the BM-MSCs isolated from mice with EAE [28].…”