Human embryonic stem cells and good manufacturing practice: report of a 1- day workshop held at stem cell biology research center, Yazd, 27[th] April 2017

This report explains briefly the minutes of a 1-day workshop entitled; "human embryonic stem cells [hESCs] and good manufacturing practice [GMP]" held by Stem Cell Biology Research Center based in Yazd Reproductive Sciences Institute at Shahid Sadoughi University of Medical Sciences, Yazd, Iran on 27[th] April 2017. In this workshop, in addition to the practical sessions, Prof. Harry D. Moore from Centre for Stem Cell Biology, University of Sheffield, UK presented the challenges and the importance of the biotechnology of clinical-grade human embryonic stem cells from first derivation to robust defined culture for therapeutic applications

comparison of two types of relaxation techniques on postoperative state anxiety in candidates for the mastectomy surgery: a randomized controlled clinical trial

Background: Anxiety among patients after surgery can affect their physiological and psychological well-being. The aim of this study was to investigate and compare the effects of Benson's relaxation and rhythmic breathing techniques on postoperative anxiety in candidates for the mastectomy surgery

Methods: This randomized controlled clinical trial study was conducted with ninety patients in 2013. The patients were hospitalized for the mastectomy surgery in three surgical wards in a teaching hospital, Tehran, Iran. They were randomly assigned into three groups: Benson's relaxation including the cognitive relaxation technique type, rhythmic breathing including the somatic relaxation technique type and control groups. According to the Davidson and Schwartz multi-process theory, the Benson's relaxation and the rhythmic breathing techniques have cognitive and somatic effects, respectively. One day before the surgery, the patients in the intervention groups were trained regarding relaxation and breathing techniques and were asked to perform the techniques under the supervision of the researcher in the night before the surgery. The cognitive somatic anxiety questionnaire was used to measure anxiety before the intervention and half an hour after recovery of consciousness after the surgery. Descriptive and inferential statistics were used for data analysis via the SPSS v.21 software

Results: There were no statistically significant differences between the groups in terms of demographic characteristics. The application of both techniques reduced the level of patients' anxiety after the surgery. The patients in the Benson's relaxation technique group reported only the relief of somatic anxiety. However, the breathing technique patients reported a reduction in both cognitive and somatic anxiety

Conclusion: The Benson's relaxation and rhythmic breathing techniques can reduce postoperative anxiety in patients after the mastectomy surgery

Relationship between sperm protamine deficiency and apoptosis in couples with unexplained repeated spontaneous abortions

Background: Etiology of more than half of Recurrent Spontaneous Abortion. The etiology of more than 50 percent of Recurrent Spontaneous Abortions [RSA] cases has been remained unexplained. It is supposed that RSA may have "paternal effect" due to supply 50% of embryonic genomic content by male gamete

Objective: The aim of present study was to evaluate the role of sperm apoptosis and protamine deficiency at same time in RSA cases

Materials and Methods: Forty fertile [control] and 40 unfertile men with RSA [case] were enrolled in this case-control study. Semen analysis was performed in accordance with WHO criteria and sperm apoptosis and protamine deficiency were evaluated by cell apoptosis detection kit and chromomycin A3, respectively

Results: Results showed significant different between normal morphology and total motility in two groups. Case group had higher percentage of spermatozoa with protamine deficiency and apoptosis compared to controls significantly

Conclusion: Our results showed that in cases of RSA, in addition to abnormal sperm parameters, we have a high percentage of spermatozoa with protamine deficiency and apoptosis and these two anomalies may consider as important causes of idiopathic recurrent abortions. It should be advised that sperm chromatin and DNA examinations are useful tools in the process of RSA treatments

Reproductive biology, stem cells biotechnology and regenerative medicine: a 1-day national symposium held at Shahid Sadoughi University of medical sciences

This paper summarizes the proceedings of a 1 day national symposium entitled "Reproductive biology, stem cells biotechnology and regenerative medicine" held at Shahid Sadoughi University of Medical Sciences, Yazd, Iran on 3[rd] March 2016. Here, we collected the papers that presented and discussed at this meeting by specialists that currently researched about the overlaps of the fields of reproductive biology and stem cells and their applications in regenerative medicine

Comparing the roles of sperm chromatin integrity and apoptosis in intrauterine insemination outcomes of couples with mild male and female factor infertility

Intrauterine insemination [IUI] is one of the therapeutic approaches for infertility. The objective of this study was to evaluate DNA integrity and apoptosis role in success of IUI in both mild male and female factor infertility. Patients were divided into two groups: M [mild male factor; n=29] and F [female factor; n=31] undergoing single IUI. Ejaculates were analyzed and chromatin quality was assessed using chromomycin A3 [CMA3] staining. In addition, spermatozoal apoptosis was recognized using TUNEL assay. Statistical analyses were done using t-test and Mann Whitney test for sperm apoptosis and sperm chromatin by SPSS. Data were expressed in mean +/- SD for variables. P<0.05 was considered statistically significant. Sperm concentration and progressive motility were higher in F than M group. Sperm with normal morphology were statistically similar in M and F infertile patients [32.7 +/- 15.6% vs. 35.5 +/- 9.07%, p=0.39]. Sperm chromatin immaturity was higher in patients with mild male infertility, when compared with the other group [p<0.01]. Also, 32.0 +/- 5.6% and 30.8 +/- 6.1% of the spermatozoa showed signs of apoptosis in groups M and F, respectively [p=0.49]. Very low [3.4%] clinical pregnancy rates were noticed in patients with mild male factor infertility. Defect in sperm motility as well as high rates of DNA damage and apoptosis may be involved with very low rate of pregnancy outcomes in patients with mild male factor infertility. Therefore, it seems the application of IUI may have better outcomes in patients with female infertility compared to mild male factor infertility

investigation on mitochondrial DNA deletions and telomere shortening during multiple passages of adult stem cells

Limited resources for adult stem cells necessitate their in vitro culture prior to Clinical use. Investigating mitochondrial DNA [mtDNA] and telomere shortening has proved to be important indications of stem cell validity. This study was designed to investigate these indicators in multiple passages of three adult stem cell lines which were produced in our stem cell laboratory. In this study, Dental Pulp Stem Cells [DPSCs], Periapical Follicle Stem Cells [PAFSCs] and Human Foreskin Fibroblast [HFF] cell lines were expanded for 20 passages. After 1, 5, 10, 15 and 20 passages, expanded cells were harvested and DNA was extracted for further studies. Common mtDNA mutation was detected by multiplex PCR and telomere shortening was tested by Southern blot analysis. The common deletion was not detected in any of the stem cells or cell lines after several passages. In addition, Southern blot analysis indicated that the mean difference of telomere length between first and last passage was 0.25 kb in DPSC, 0.1 kb in PAFSC and 0.32 kb in HFF which indicates that the mean telomere length in various passages of the samples showed insignificant changes. Absence of mtDNA mutations in adult stem cell lines indicates good mitochondrial function even after 20 passages. In addition, absence of telomere shortening indicates stem cells validity after multiple passages. It is hoped this information could pave the way for using in vitro expansion of adult stem cells for future Clinical applications

Mesenchymal Stem Cells Improved the Ultrastructural Morphology of Cerebral Tissues after Subarachnoid Hemorrhage in Rats

Subarachnoid hemorrhage (SAH) causes widespread disruption in the cerebral architecture.The process of SAH is complicated and many people lose their lives or become disabled after injury. Mesenchymal stem cells (MSCs) are considered as good candidate for repair of cerebral damage. The aim was to assess the ultrastructural changes in the rat cerebral tissue after intravenous transplantation of MSCs. Female Wistar rats (8 per group) weighing 275~300 g were assigned to control (SAH+PBS) and experimental groups (SAH+MSCs).The samples from middle cerebral arterial wall and parietal cerebral tissue were prepared for transmission electron microscopy (TEM) according to standard protocol. Fine architectures of the vessel wall, including the contraction of the inner layer, smooth muscle layer,as well as neural cells were observed after SAH. Cerebral arterial wall and cortex, including neuronal and glial cells were injured post SAH. But, administration of MSCs improved the structural integrity of cerebral tissues. Changes were much more balanced with their relative improvement in some areas. The role of MSCs for repairing the injured cerebral tissues post experimental SAH was approved by electron microscopy.

Characterization of mesenchymal stem cells from human dental pulp, preapical follicle and periodontal ligament

Human dental stem cells have high proliferative potential for self-renewal that is important to the regenerative capacity of the tissue. The aim was to isolate human dental pulp stem cells [DPSC], periodontal ligament stem cells [PDLSC] and periapical follicle stem cells [PAFSC] for their potential role in tissue regeneration. In this experimental study, the postnatal stem cells were isolated from dental pulp, preapical follicle and periodontal ligament .The cells were stained for different stem cell markers by immunocytochemistry. To investigate the mesenchymal nature of cells, differentiation potential along osteoblastic and adipogenic lineages and gene expression profile were performed. For proliferation potential assay, Brdu staining and growth curve tests were performed. Finally, all three cell types were compared together regarding their proliferation, differentiation and displaying phenotype. The isolated cell populations have similar fibroblastic like morphology and expressed all examined cell surface molecule markers. These cells were capable of differentiating into osteocyte with different capability and adipocyte with the same rate. PAFSCs showed more significant proliferation rate than others. Reverse transcriptase PCR [RT-PCR] for nanog, oct4, Alkaline phosphatase [ALP] and glyceraldehydes-3-phosphate dehydrogenease [GADPH] as control gene showed strong positive expression of these genes in all three isolated cell types. PDLSCs, DPSCs and PAFSCs exist in various tissues of the teeth and can use as a source of mesenchymal stem cells for developing bioengineered organs and also in craniomaxillofacial reconstruction with varying efficiency in differentiation and proliferation