Fibrin hydrogel as a scaffold for differentiation of induced pluripotent stem cells into oligodendrocytes.

The importance of tissue engineering has been established as a promising approach in treating neurodegenerative diseases. The purpose of the current study is to determine the effect of fibrin hydrogel on the differentiation of iPSC into oligodendrocyte. For this purpose, iPSCs transduced by miR-338 expressing lentiviruses. They were treated with basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and platelet-derived growth factor (PDGF)-AA. The process was traced by a 6-day treatment in a mitogen-free medium. At the end of the process, multipolar preoligodendrocytes appeared. In comparison to tissue culture plate (TCP), MTT assay demonstrated a significant increase in the viability of cells cultured in fibrin hydrogel. SEM analysis showed cells with elongated morphology and intertwined intercellular interactions. An immunofluorescent assay confirmed the expression of oligodendrocyte markers Olig2 and O4. In comparison to TCP, real-time PCR data indicated a significant increase in the expression of some markers such as Olig2, MBP, Sox10, and PDGFRα on cells encapsulated in fibrin hydrogel. Overall, the results suggest that fibrin hydrogel improves viability of cells and promotes the differentiation of iPSCs into preoligodendrocytes. Hence, it can be used as an appropriate option in the tissue engineering in order to treat neurodegenerative diseases. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:192-200, 2020.

Human olfactory stem cells: As a promising source of dopaminergic neuron-like cells for treatment of Parkinson's disease.

The production of dopaminergic (DA) neurons from stem cells holds a great promise for future clinical treatment of neurodegenerative diseases, such as Parkinson's disease (PD). Olfactory ecto-mesenchymal stem cells (OE-MSCs) derived from the adult human olfactory mucosa can be easily isolated and expanded in culture while maintaining their immense plasticity. In addition to reduced ethical concerns, OE-MSCs could be transplanted across allogeneic barriers, making them desirable stem cells for clinical applications. The goal of this study was to define the potentiality of human olfactory mucosa-derived MSCs aimed at differentiation into DA neuron-like cells. OE-MSCs were induced to differentiate to DA neuron-like cells in vitro by using sonic hedgehog (SHH), fibroblast growth factor 8 (FGF8), basic fibroblast growth factor (bFGF), Glial cell line-derived neurotrophic factor (GDNF) and brain derived neurotrophic factor (BDNF). Then the differentiated neurons were characterized for expression of DA neuron markers by Real-time PCR, immunocytochemistry and flow cytometry. Our findings showed that differentiated OE-MSCs could significantly express DA neuron markers at mRNA and protein levels along with dopamine release 12 days post-differentiation. These results support the viability and feasibility of using OE-MSCs as a source of in vitro generated DA neuron-like cells for treatment of DA disorders namely PD.

The effect of purmorphamine on differentiation of endometrial stem cells into osteoblast-like cells on collagen/hydroxyapatite scaffolds.

We assessed the effect of purmorphamine along with collagen/hydroxyapatite scaffold in inducing osteogenesis of human endometrial stem cells (hEnSCs). The adhesion, viability, proliferation, and differentiation of cells on scaffold were assayed with SEM, MTT, real time-PCR, and ALP assay, respectively. The results were shown good integration of cells with scaffold. Also, qRT-PCR of differentiated cells shows that osteoblast cell markers are expressed after 21d in 2D and scaffold groups while in the scaffold group the expression of these markers were higher than the 2D group. Based on our findings, collagen/hydroxyapatite scaffold with PMA has the potential role in osteogenic differentiation of hEnSCs.

Differentiation of Human Endometrial Stem Cells into Schwann Cells in Fibrin Hydrogel as 3D Culture.

Human endometrial stem cells (hEnSCs) are a new source of adult multipotent stem cells with the ability of differentiation into many cell lineages. Many stem cell sources are desirable for differentiation into Schwann cells. Schwann-like cells derived from hEnSCs may be one of the ideal alternative cell sources for Schwann cell generation. In this study, for differentiation of hEnSCs into Schwann cells, hEnSCs were induced with RA/FSK/PDGF-AA/HRG as an induction medium for 14 days. The cells were cultured in a tissue culture plate (TCP) and fibrin gel matrix. The viability of cultured cells in the fibrin gel and TCP was analyzed with 3-[4,5-dimethyl-2-thia-zolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay for 7 days. The attachment of cells was analyzed with SEM and DAPI staining. The expression of S100 and P75 as Schwann cell markers was evaluated by immunocytochemistry and quantitative real-time PCR (RT-PCR). The evaluation of the MTT assay and gene expression showed that the survival rate and differentiation of hEnSCs into Schwann cells in the fibrin gel were better than those in the TCP group. These results suggest that human EnSCs can be differentiated into Schwann cells in the fibrin gel better than in the TCP, and the fibrin gel might provide a suitable three-dimensional (3D) scaffold for clinical applications for cell therapy of the nervous system.