Another drawback is low engraftment due to immunological rejection of the transplanted cells by the host immune system, although it was shown that short-term treatments with immunosuppressive drugs in mice enhanced the engraftment of unrelated pluripotent stem cells [59]

Another drawback is low engraftment due to immunological rejection of the transplanted cells by the host immune system, although it was shown that short-term treatments with immunosuppressive drugs in mice enhanced the engraftment of unrelated pluripotent stem cells [59]. Alternatively, MSCs are generally regarded as a safe, non-tumorigenic and low-immunogenic adult stem cell type (with several clinical trials ongoing) [60], that seem to have the potential to differentiate into neural-like cells [7], [9], hence could represent an interesting alternative PRKM3 to pluripotent stem cells for clinical use. as mesenchymal stromal cells, are defined as multipotent adult stem cells, possessing self-renewal capacity and multilineage differentiation potential [1], [2]. MSCs were originally identified in the bone marrow [3], but more recently, cells with characteristics similar to MSCs have been identified in many other locations, such as perivascular regions of multiple organs and tissues (like the fat tissue) [4] and several regions of the umbilical cord, namely the umbilical cord matrix (also known as the Wharton’s jelly) [5]. MSCs have been characterized as a safe, available, Alfuzosin HCl low-immonogenic and clinically promising adult stem cell type [1], [5], [6]. Several reports in the literature have shown the potential of MSCs to differentiate into neural stem-like cells [7]C[9]. Despite controversy about MSCs (a mesenchymal cell type) differentiating into neural-like cellular fates, compelling evidence has shown that indeed MSCs express neuroectodermal markers, like nestin [8], [10]C[13] and have at least a partial neural crest, neuroepithelial origin [14], [15], suggesting plasticity towards neural-like lineages, opening research avenues for the treatment of distinct neurodegenerative diseases [16], [17]. MSCs have been rather explored in terms of neuronal-like differentiation [8], [13], [18]C[20], but the Alfuzosin HCl first reports addressing oligodendrocyte-like specification were only published recently [21], [22]. Nevertheless, further studies are required to fully address this potential. Demyelination of the central nervous Alfuzosin HCl system (CNS) is caused by loss of oligodendrocytes (OLs) and may occur as a result of traumatic injury or non-traumatic neurodegenerative diseases, like multiple sclerosis (MS). Remyelination of the affected areas is typically low and demyelinated areas become inflamed and populated by astrocytes, causing the formation of scar tissue [23]. Stem cell-based approaches that allow for a quicker and more robust remyelination of the affected areas are considered promising for the treatment of demyelinating diseases. However, despite recent advances regarding oligodendroglial differentiation of pluripotent stem cells (namely human embryonic stem cells – hESCs [24], [25] and induced pluripotent stem cells – iPSCs [26]), these are not yet considered safe for application in a clinical setting. Hence, the current lack of appropriate and safe cell sources hamper the use of stem cell-based approaches for the treatment of demyelinating diseases in the clinic. The objectives of the present work were to thoroughly characterize human MSCs isolated from the umbilical cord matrix (UCM) and assess whether these cells possessed neural- and more specifically, oligodendroglial-like differentiation capacity. The results presented here suggest that umbilical cord matrix mesenchymal stem cells (UCM-MSCs) possess a certain degree of plasticity to differentiate into neural-like cells, and subsequently into cells with phenotypic characteristics of oligodendrocyte precursors and immature oligodendrocytes. Despite the need for testing further differentiation protocols and to perform functional studies to assess the full potential of these cells, the results presented here are promising in the context of cell-based therapeutic strategies for demyelinating diseases. Materials and Methods Isolation and culture of human mesenchymal stem cells (MSCs) from the umbilical cord matrix (UCM) Human umbilical cords were obtained after birth from healthy donors, with written informed consent of the parent(s) and the study was approved by the Ethics Committee of Alfuzosin HCl Maternidade de Bissaya Barreto C Centro Hospitalar de Coimbra (ref. 356/Sec). Samples were stored at room temperature (RT) in sterile 50 ml conical tubes (VWR International) for 12 to 48 h before tissue processing. The isolation procedure of MSCs was adapted from a protocol described by Reinisch the population doubling (PD), as described [28]. The PD for each passage was calculated and added to the PD of the previous passages to generate data for cumulative population doublings (CPD). In addition, the generation time (GT) – average time between two cell doublings – was calculated from P2 to P8 using the following formula, as described [29]: ?=? [log10(2) x ?=? x B/and undifferentiated MSCs were used as the control sample. The PCR cycling parameters were 94C for 5 min; 30 cycles.