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Microtubule Stabilizer Epothilone B As a Motor Neuron Differentiation Agent for Human Endometrial Stem Cells Publisher Pubmed



Mahmoodi N1 ; Ai J2 ; Ebrahimibarough S2 ; Hassannejad Z3 ; Hasanzadeh E4 ; Basiri A2 ; Vaccaro AR5 ; Rahimimovaghar V1
Authors
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Authors Affiliations
  1. 1. Sina Trauma and Surgery Research Center, Sina Hospital, Tehran University of Medical Sciences, Hasan-Abad Square, Imam Khomeini Ave., Tehran, 11365-3876, Iran
  2. 2. Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Number 88, Italy Street, Between Ghods Street and Vesal Shirazi Street, Tehran, 14177-55469, Iran
  3. 3. Pediatric Urology and Regenerative Medicine Research Center, Children's Medical Center, Pediatric Center of Excellence, Tehran University of Medical Sciences, No. 62, Dr. Gharibs Street, Keshavarz Boulevard, Tehran, 1419733151, Iran
  4. 4. Department of Tissue Engineering, School of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Next to Tooba Medical Building, Khazar Boulevard, Sari, 48471-91971, Iran
  5. 5. Department of Orthopedic Surgery, Rothman Institute, Thomas Jefferson University, 1925 Chestnut Street, 5th Floor, Philadelphia, 19107, PA, United States

Source: Cell Biology International Published:2020


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. © 2020 International Federation for Cell Biology
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