Author/Authors :
Hosseini, Ali Blood Transfusion Research Center - High Institute for Research and Education in Transfusion Medicine - Tehran, Iran , Amiri, Fatemeh Department of Medical Laboratory Science - Paramedicine Faculty - Hamadan University of Medical Science - Hamadan, Iran , Khalighi, Fereshteh School of Veterinary Science - Shiraz, Iran , Mohammadi Roushandeh, Amaneh Department of Medical Biotechnology - Paramedicine Faculty - Guilan University of Medical Sciences - Rasht, Iran , Habibi Roudkenar, Mehryar Department of Medical Biotechnology - Paramedicine Faculty - Guilan University of Medical Sciences - Rasht, Iran , Kuwahara, Yoshikazu Faculty of Medicine - Tohoku Medical and Pharmaceutical University - Sendai, Japan , Bashiri, Hamed Department of Medical Laboratory Sciences - faculty of Paramedical - Kurdistan University of Medical Sciences - Sanandaj, Iran
Abstract :
Due to oxidative stress, hypoxia, and serum
deprivation, a large percentage of mesenchymal stem cells
(MSCs) die in the early stages of transplantation. The present
study aimed to address whether induction or inhibition of
autophagy would affect the viability of MSCs after exposure to
oxidative stress.
Methods: MSCs were isolated from umbilical cord tissue using
the Ficoll gradient method. pCMV-GFP-LC-3 plasmid containing
GFP-tagged LC3 was transfected into MSCs to assay autophagy
level in these valuable cells. The four study groups were: MSCLC3-
Rapa, MSC-LC3-3MA, MSCs without any transfection,
and MSC-GFP-LC3 (control groups). To induce autophagy, the
MSC-GFP-LC3 was treated with different concentrations of Rapa
for 24 hours and named MSC-LC3-Rapa. To inhibit autophagy in
MSC-GFP-LC3, these cells were cultured in the presence of 3MA
for 24 hours and named MSC-LC3-3MA. Non-treated MSCGFP-
LC3 and MSCs were considered as control groups. MSCs
were exposed to lethal doses of H2O2 followed by cell viability
evaluation with the water-soluble tetrazolium salt assay method.
The data were analyzed with SPSS version 18.0 using one-way
ANOVA test. P<0.05 was considered statistically significant.
Results: The results revealed that the enhancement of autophagy
in MSC-LC3-Rapa sensitized them against oxidative stress
(P=0.0006) and inhibition of autophagy in MSC-LC3-3MA led
to resistance against oxidative stress (P=0.0003).
Conclusion: Inhibition of autophagy, as a non-genetic engineering
method, in MSCs enhances cell viability following exposure to
the oxidative stress. This may provide a novel strategy to promote
the efficiency of MSC-based cell therapy for clinical applications.
Keywords :
3-methyladenine , Oxidative stress Sirolimus , Autophagy , Mesenchymal stromal cells
