Publications by authors named "Afshin Pirnia"

6 Publications

  • Page 1 of 1

A comparison of the effects of fetal bovine serum and newborn calf serum on cell growth and maintenance of cryopreserved mouse spermatogonial stem cells.

Mol Biol Rep 2020 Dec 19;47(12):9609-9614. Epub 2020 Nov 19.

Medical Technology Research Center, Institute of Health Technology, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Serum is a common supplement that is widely used to protect various cells and tissues from cryopreservation because it provides the necessary active components for cell growth and maintenance. In this study, we compared the effects of newborn calf serum (NCS) and fetal bovine serum (FBS) on the cryopreservation of mouse spermatogonial stem cells (SSCs). The isolated SSCs were cryopreserved in two groups: freezing medium that contained 10% DMSO (dimethyl sulfoxide) and 10% FBS in DMEM (Dulbecco's Modified Eagle's Medium) (group 1) and freezing medium that contained 10% DMSO and 10% NCS in DMEM (group 2). Real-time PCR was performed for stemness gene expression. The SSCs' viability was performed by trypan blue. We observed that the SSCs had increased viability in the NCS-freeze/thaw group (87.82%) compared to the FBS-freeze/thaw group (79.83%), but this increase was not statistically significant (P < 0.105). Promyelocytic leukemia zinc finger (Plzf) and Lin28 gene expression levels in the NCS-frozen/thawed SSCs were not significantly different compared to the FBS-frozen/thawed SSCs; however, Nanog gene expression increased considerably, and Dazl gene expression decreased significantly. The results in this study demonstrated that the presence of NCS in a solution of cryopreserved SSCs increased their viability after freeze/thawing and might promote the proliferation of cultivated SSCs in vitro by increasing the relative expression of Nanog.
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http://dx.doi.org/10.1007/s11033-020-06004-2DOI Listing
December 2020

The role of vasopressin V1A and oxytocin OTR receptors in protective effects of arginine vasopressin against HO-induced oxidative stress in H9C2 cells.

Arch Physiol Biochem 2020 Mar 6:1-6. Epub 2020 Mar 6.

Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences Khorramabad, Iran.

Oxidative stress, has been shown to play an important role in the pathophysiology of cardiac remodelling and heart failure. The aim of study is effect of arginine vasopressin (AVP) on apoptosis of cardiomyocyte via its receptors. The cell viability effect of AVP in H9C2 cardiomyocytes was assayed using the MTT method. The transcription and translation level of apoptosis genes (Bax, Bcl-2, caspase-3) were discovered with qRT-PCR and western blotting. The results showed that vasopressin could reduce apoptosis in cardiomyocytes cell line through downregulation of caspase-3, BAX and upregulation of Bcl-2 ( < .001). Also, there was a decrease in anti-apoptosis effect of vasopressin when V1A and OTR receptors were blocked with their antagonists. These results suggest that activation of V1A and OTR receptors in H9C2 cells mediate protective effect of vasopressin via regulating apoptosis marker that lead to cell survival under conditions of stress oxidative.Key pointAVP may contribute to the improvement of heart ischaemia through its actions on V1A and OTR receptors.
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http://dx.doi.org/10.1080/13813455.2020.1729816DOI Listing
March 2020

Effect of selenium on freezing-thawing damage of mice spermatogonial stem cell: a model to preserve fertility in childhood cancers.

Stem Cell Investig 2019 26;6:36. Epub 2019 Nov 26.

Department of Anatomical Sciences, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Background: During treatment of childhood cancers, fertility of boys may be affected. Therefore, freezing spermatogonial stem cell (SSC) is recommended. However, freezing-thawing process may cause damage to SSCs. This study was conducted to evaluate protective effects of selenium on freezing-thawing damage of mice SSCs using investigation of cell viability and investigation of apoptosis related genes expression including , , , and .

Methods: SSCs were extracted from 80 6-day-old mice. The SSCs were divided into four groups: cryopreservation along with selenium (low and high dose), vitrification along with selenium (low and high dose), cryopreservation control, and vitrification control. Trypan blue staining and real-time polymerase chain reaction (real-time PCR) were used to investigate cell viability and gene expression, respectively.

Result: Comparison of cell viability in the experimental groups did not show a significant association. Expression of and was significantly lower in cryopreservation group with low-dose selenium. Expression of was significantly lower in cryopreservation group with high-dose selenium. Expression of and was significantly lower in vitrification group with low-dose selenium, and expression of was significantly upper. Expression of and was significantly lower in vitrification group with high-dose selenium, and expression of was significantly upper (P<0.001).

Conclusions: Selenium had dose dependent effect on apoptosis related genes profile. The only evident effect was the effect of low-dose selenium in cryopreservation on inhibition of apoptosis via extrinsic pathway.
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http://dx.doi.org/10.21037/sci.2019.10.01DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6917556PMC
November 2019

The Effect of Caffeic Acid on Spermatogonial Stem Cell-type A Cryopreservation.

Rep Biochem Mol Biol 2018 Oct;7(1):85-93

Razi Herbal Medicine Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran.

Background: Cancer treatment methods can lead to male infertility .in this regard, cryopreservation of spermatogonial stem cells (SSC) and cell-to-person transplantation after the course of treatment to resolve the problem of infertility is a good one. The cryopreservation of SSC is an important process as it can help on the return of spermatogenesis. However, during this process, the stem cells often become damaged which degrades their value for experiments and treatments. Caffeic acid (CA) is an antioxidant that has been shown to increase the viability of cells under stress. The aim of this study was to investigate the effect of CA has on spermatogonial stem cell (SSC) cryopreservation.

Methods: Spermatogonial stem cells isolated from the testes of Balb/c mice pups were cultured in laminincoated dishes, purified using CD90.1 microbeads, then cryopreserved in vitrification media supplemented with 10 µM CA either through a slow or rapid freezing process. After thawing, cell viability was evaluated. Expression of Bax, Fas, Bcl-2 and P53 genes was determined by real-time PCR. Gel electrophoresis was used to confirm the results of the real-time PCR.

Results: The viability of the SSCs that were rapidly frozen and treated with CA was observed to be significantly reduced compared to the control group (p < 0.003). The viability SSCs that received CA and underwent the slow freezing treatment was significantly reduced compared to controls (p < 0.002). The expression levels of BAX, BCL-2, and Fas in the rapid freeze-thaw group didn't significantly change. However, the levels of P53 expression were shown to increase. In the group of SSCs that underwent the slow freezing process, the BAX gene expression levels increased, while the levels of BCL-2 gene expression decreased. No significant changes in the level of Fas and P53 expression were detected. When comparing the groups that received CA treatment, SSCs that were rapidly frozen showed an up-regulation of Fas and P53 expression and a down-regulation of Bcl-2 and Bax expression.

Conclusion: Caffeic acid may protect intact SCCs during the cryopreservation process through stimulating the induction of apoptosis in injured SSCs. Supplementing the vitrification media with CA has a superior effect on the preservation of SSCs.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6175585PMC
October 2018

Evaluation of alginate hydrogel cytotoxicity on three-dimensional culture of type A spermatogonial stem cells.

Int J Biol Macromol 2017 Feb 27;95:888-894. Epub 2016 Oct 27.

Razi Herbal Medicine Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran; Department of Anatomical Sciences, Lorestan University of Medical Sciences, Khorramabad, Iran. Electronic address:

The culture of spermatogonial cells for future transplantation, based on the specific biology of these cells is important and necessary. Recently, the use of scaffolds especially alginate for culturing stem cells has been the focus of many researchers. The aim of this study was to evaluate the cytotoxicity of alginate hydrogels to cultures of type A spermatogonial stem cells. Spermatogonial stem cells of 6day-old immature mice were isolated by surgery; thereafter, the cells were purified by MACS using antibodies against thy-1 and C-kit and cultured on a layer of laminin. After purification, spermatogonial stem cells were encapsulated in alginate hydrogels. After one month of encapsulation and culture in DMEM culture medium containing 10ng/ml GDNF, cells were removed from hydrogel and were examined for viability, cell morphology and structure, cytotoxicity and expression of apoptosis genes Fas, P53, Bax, Bcl2, Caspase3 by staining with trypan blue, scanning electron microscopy, LDH test, and Real time PCR, respectively. The encapsulation did not change the morphology and viability of spermatogonial stem cells. Investigations showed that spermatogonial stem cells preserve by the high viability (74.08%) and cytotoxicity of alginate hydrogel was estimated to be 5%. Expression of Fas gene increased in main group compared with the control group, and expression of Bax and P53 was reduced in main group compared with the control group. Expression of Bcl2 and Caspase3 genes did not show any significant difference between the main group and the control group. Considering the lack of cytotoxicity and antioxidant properties of alginate hydrogel scaffold and high viability of cells, this three-dimensional scaffold is applicable for culturing and encapsulation of spermatogonial stem cells.
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http://dx.doi.org/10.1016/j.ijbiomac.2016.10.074DOI Listing
February 2017

Supplementation freeze-thawed media with selenium protect adipose-derived mesenchymal stem cells from freeze-thawed induced injury.

Cryobiology 2016 10 19;73(2):135-9. Epub 2016 Aug 19.

Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran; Department of Anatomical Sciences, Lorestan University of Medical Science, Khorramabad, Iran. Electronic address:

Successful freezed-thaw of adipose-derived mesenchymal stem cells (ADMSCs) could be a major step in regenerative medicine as well as in the cloning of animal breeds. The aim of this study was to evaluate the efficacy of selenium on the optimizing of freezed-thaw media in the ADMSCs. ADMSCs were extracted from NMRI mice and purified with positive selection Monoclonal CD105 Antibody (PE) and negative selection Monoclonal CD31 and CD45 Antibody using MACS method as well as differentiation to adipose and bone tissue. ADMSCs were divided into four groups. ADMSCs were freezed-thaw under standard condition with or without the addition of 5 ng/ml selenium to both the cryopreservation and thawing solutions. Frozen cells were thawed after four months and viability and cytotoxicity of the cells were analyzed by the Trypan blue test and MTT assay respectively. RNA was extracted and cDNA was synthesized and the expression of apoptotic genes (P53, Fas, Bax, Caspase3, and Bcl2) was examined using Real time-PCR Rotor gene 2009. This study compares slow and rapid methods of cryopreservation. After thawing, viability of the cells treated with selenium was higher than the control group in rapid and slow cryopreserved ADMSCs. Also, the percentage of living cells in the slow cooling method was considerably more than with the rapid cooling method. After analysis of the results using Real time-PCR, the Bcl2 gene was shown to be expressed in both the rapid and slow cooling methods. In the rapid cooling group in addition to the BCL-2 gene, p53 was also expressed. It appears that selenium prevented the apoptotic genes from expression due to its anti-apoptotic effects. The slow cooling method is better and more optimized for ADMSCs protecting them from oxidative damage to a greater extent compared to the rapid cooling method.
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http://dx.doi.org/10.1016/j.cryobiol.2016.08.009DOI Listing
October 2016