Microtubule stabilizer epothilone B as a motor neuron differentiation agent for human endometrial stem cells

Abstract

Microtubule-stabilizing agents (MSAs), until now, have primarily been considered for their anti-proliferative effects in the setting of cancer. However, recent studies have revealed that one particular MSA, epothilone B (EpoB), can promote axonal regeneration after traumatic spinal cord injuries (SCI) even in the presence of inhibitor molecules such as neurite outgrowth inhibitor-A (Nogo-A). On the basis of the importance of having an efficient motor neuron (MN) differentiation protocol for stem cell therapy and the attention of MSAs for SCI treatment, our study investigated the effect of EpoB on human endometrial stem cells (hEnSCs) differentiation into MN-like cells. hEnSCs were isolated and characterized by flow cytometry. The hEnSC cell viability was evaluated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To mimic the in vivo inhibitory environment, hEnSCs were also differentiated in the presence of Nogo-A. After 15 days of differentiation, the expressions of MN-markers were evaluated by real-time reverse-transcriptase polymerase chain reaction and immunofluorescence. According to the MTT assay results, three doses (1, 5, and 10 nM) of EpoB were selected to evaluate their effect on MN-differentiation. All selected doses can increase the efficacy of hEnSCs differentiation into MN-like cells. In particular, the 10 nM EpoB dosage was shown to increase the axon elongation, cell alignment, and upregulation of these MN-markers compared with other doses. EpoB can improve MN differentiation from hEnSC and potentially provide a unique route for neuronal replacement in the setting of SCI.

Tissue‐engineered nerve graft using silk‐fibroin/polycaprolactone fibrous mats decorated with bioactive cerium oxide nanoparticles

“…The mean porosity and pore size of fabricated neural guidance conduit (NGC) was 69 ± 4.5% and 11.1 ± 2.1 nm which proved this NGC could be utilized as a promising strategy to bridge the gap between severed peripheral nerve stumps for the treatment of damaged nerves that the results confirmed designed nerve conduit fabricated by electrospinning with tailored mechanical strength, hydrophilic/hydrophobic property and desirable cell adhesion properties could be an efficient alternative to autograft for peripheral nerve regeneration research. Due to its advantage of high‐surface area and existence of bioactive inorganic ions respectively for cell attachment and tissue regeneration, it is believed that this electrospun nerve conduit could find more application on behalf of nerve regeneration in future, to further improve functional regeneration outcome especially for longer nerve defect restoration 7‐10,41 …”

Proanthocyanidin as a crosslinking agent for fibrin, collagen hydrogels and their composites with decellularized Wharton’s-jelly-extract for tissue engineering applications

“…Lentivirus (second-generation) was utilized for GFP transduction of hEnSCs, based on a previous study. 29 For this purpose, plasmid pCDH-Cop GFP-Pru was used comprising EF-1α promotor to upgrade the GFP transgene expression. HEK293 T cell line was cultured and transfected with the vector linked with a pMD2.G as an envelope plasmid and psPAX2 packaging plasmid using the calcium phosphate precipitation method.…”

Defining the role of 17β‐estradiol in human endometrial stem cells differentiation into neuron‐like cells

“…Up to now, there are no effective treatments for full functional retrieval after an SCI for the limited regeneration ability of the CNS. One potential solution is the development of stem cell differentiation to substitute damaged and lost cells following SCI (Mahmoodi et al, 2020). Recent studies have shown that the use of predifferentiation stem cells before transplantation can reduce the risk of stem cell proliferation or their migration to inappropriate areas of the CNS that may lead to severe dysfunction or F I G U R E 8 The results of quantitative real-time PCR analysis on day 14 for neural markers in hEnSCs after incubation with the two utilized neuronal differentiation media (with and without 17β-estradiol).…”