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Showing 5 results for Pasbakhsh

Reza Mahmoudi, Aligholi Subhani, Parichehr Pasbakhsh, Farideh Etesam, Iraj Amiri, Mozhdeh Salehnia, Farid Abolhasani,
Volume 3, Issue 2 (7-2005)

Background: In vitro maturation (IVM) of oocytes is a promising technique to reduce the costs and avert the side-effects of gonadotropin stimulation for in vitro fertilization (IVF). The pregnancy rates from oocytes matured in vitro are much lower than those of in vivo stimulation cycles, indicating that optimization of IVM remains a challenge. Objective: In this study, we investigated the effect of cumulus cells on maturation and fertilization rate of immature oocytes (Germinal vesicle). Materials and Methods: Germinal vesicle (GV) oocytes were recovered from 6-8 weeks old Balb C female mice 48hr after injection of 10 IU pregnant mare serum gonadotropin (PMSG). Collected oocytes were divided into two groups. Group A: GV oocytes without cumulus (denuded oocyte). Group B: GV oocytes with cumulus cells (cumulus-oocyte complex). The oocytes in both groups were cultured in TCM-199 medium in a humidified atmosphere of 5% CO2 in air at 37�C. The maturation, fertilization and developmental rates were recorded after 24hr. Results: Maturation, fertilization and developmental rates in denuded oocytes (DO) were 65.1%, 68.02%, 78.63% respectively, and in cumulus-oocyte complex (COC) were 78.20%, 85.57% and 85.05%, respectively. The maturation, fertilization and developmental rates of COC were significantly higher than those of DO (p<0.05). Conclusion: The results show that cumulus cells have beneficial effects on maturation, fertilization and cleavage rates of mice oocytes.
Mitra Bakhtiari, Aligholi Sobhani, Mohammad Akbari, Parichehr Pasbakhsh, Mehdi Abbasi, Azim Hedayatpoor, Fardin Amidi , Feridoon Sargolzaei,
Volume 5, Issue 3 (7-2007)

Background: Various approaches have been used in the attempts to improve the quality of frozen–thawed mouse sperms. According to literatures, it seems that hyaluronic acid (HA) has an important role on the permeability and motility of sperms and their interaction with gametes.
Objective: For evaluation of HA supplementation on sperm characteristics and fertilization capability, we investigated the effect of different doses of HA on mouse sperm morphology, motility, vitality and fertilization capability after freezing and thawing.
Materials and Methods: The cauda epididymes was removed from 6 male mice with aseptic method. The sperm samples were frozen in 1.8 ml cryotubes with 18% raffinose and 3% skimmed milk containing cryo-protectant solution. HA at the concentration of 750, 1000 or 1250 µg/ml was supplemented to frozen-thawed sperms. Sperm motility was measured with microscope, and fertilization rate was evaluated after routine IVF by counting the fertilized oocytes. For sperm morphology, papaniclau staining was used while; Eosin B was used for the assessment of sperm viability rate.
Results: HA supplementation (750 µg/ml) improved motility parameters (p < 0.05) and increased the fertility rate (p < 0.05). The effect of 1,000 µg/ml HA was also positive on the sperms. But 1,250 µg/ml HA had negative effect on above mentioned characteristic. On the other hand, none of these doses had any effect on sperm morphology.
Conclusion: The dose of 750 µg/ml of HA has the greatest effect on the motility, vitality and fertility rate of sperms after cryopreservation.

Amaneh Mohammadi Roushandeh, Parichehr Pasbakhsh, Zohreh Alizadeh, Mehryar Habibi Roudkenar,
Volume 5, Issue 5 (7-2007)

Background: Preparation of oocytes is one of the critical factors that determine the developmental competence of embryos produced by in vitro fertilization (IVF).
Objective: In this study, the effect of cysteamine, type of media and glutathione (GSH) level on blastocysts development after in vitro maturation of mouse oocytes were investigated.
Materials and Methods: Premature female mice were primed with pregnant mare stimulating gonadotrophin (PMSG), and germinal vesicle (GV) stage oocytes were obtained 45 hr later. GV oocytes were cultured in presence of 0, 50, 100, 200 and 500 µm cysteamine in TCM199 and MEME media. After IVM, MII oocytes were in vitro fertilized (IVF) and in vitro cultured (IVC) in order to observe embryo development. A group of In Vivo Ovulated (IVO) oocytes after priming with PMSG and HCG also were included in this study. 5,5-Dithio-bis (2nitrobenzoic acid) DTNB-recycling protocol was used for GSH assay.
Results: Rate of IVM and IVF were improved in all oocytes treated with cysteamine in the two medium except 500 µm (81% MII rate in TCM and 64% MII in MEME). Rate of blastocyst in 100 µm cysteamine in TCM1199 and 200 µm in MEME was higher compared to control groups (In TCM 45% and in MEME 35%). In vivo MII and GV oocytes represented the highest and lowest GSH level respectively.
Conclusion: Our results revealed that the media and concentration of cysteamine can affects on IVM, IVF and rate of blastocysts development on dose dependant manner.
Reza Mahmoudi, Iraj Amiri, Parichehr Pasbakhsh, Iraj Ragardi Kashani, Mehdi Abbasi, Farid Aboulhasani, Tooba Mehrannia, Aligholi Sobhani,
Volume 6, Issue 5 (7-2008)

Background: Routine oocytes cryopreservation remained an elusive technique in the wide ranges of available assisted reproductive technologies. The microtubules of oocytes are vulnerable to cryoprotectants and thermal change during cryopreservation.
Objective: The effects of a vitrification protocol were investigated on the spindle and chromosome configurations of mice oocytes cryopreserved at the germinal vesicle stage.
Materials and Methods: Germinal vesicle with cumulus cells were transferred to vitrification solution which was composed of 30% (v/v) ethylene glycol, 18% Ficoll-70 and 0.3 M Sucrose either by single step or in step-wise way. Following vitrification and in vitro maturation (MII), the matured oocytes were immonostained for meiotic spindles and chromosomes, before visualization using fluorescent microscopy.
Results: A statistically significant increase was observed in the survival and maturation rate in step-wise vitrification (88.96% and 71.23% respectively) compared to single step vitrification (70.6% and 62.42% respectively) (p<0.05). Normal spindle morphology after vitrification-thawing in step-wise vitrification group (77.26%) was higher than single step vitrification group (64.24%) but lower than control group (94.75%) (p<0.05).
Conclusion: The results suggest that vitrification with step-wise procedure on mice germinal vesicle oocytes has positive effects on survival and maturation rate and normal spindle configuration compare with single step vitrification procedure.
Maryam Hosseinzadeh Shirzeily, Parichehr Pasbakhsh, Fardin Amidi, Kobra Mehrannia, Aligholi Sobhani ,
Volume 11, Issue 12 (1-2013)

Background: Recent publications about differentiation of stem cells to germ cells have motivated researchers to make new approaches to infertility. In vitro production of germ cells improves understanding differentiation process of male and female germ cells. Due to the problem of using embryonic stem cells (ESC), it’s necessary the mentioned cells be replaced with some adult multi-potent stem cells in laboratories.
Objective: The aim of this study was to obtain germ cells from appropriate source beyond ESC and compare differential potentials of adipocytes derived stem cells (ADMSCs) with bone marrow derived stem cells (BMMSCs).
Materials and Methods: To find multi-potential entity, after providing purified ADMSCs and BMMSCs, differentiation to osteoblast and adipocyte was confirmed by using appropriate culture medium. To confirm mesenchymal lineage production superficial markers (expression of CD90 and CD44 and non-expression of CD45 and CD31) were investigated by flowcytometry. Then the cells were differentiated to germ cells in inductive medium containing retinoic acid for 7days. To evaluate germ cells characteristic markers [Dazl (Deleted in azoospermia-like), Mvh (Mouse vasa homolog gene), Stra8 (Stimulated by retinoic acid) and Scp3 (Synaptonemal complex protein 3)] flowcytometry, imunoflorescence and real time PCR were used.
Results: Both types of cells were able to differentiate into osteoblast and adipocyte cells and presentation of stem cell superficial markers (CD90, CD44) and absence of endothelial and blood cell markers (CD31, CD45) were confirmative The flowcytometry, imunoflorescence and real time PCR results showed remarkable expression of germ cells characteristic markers (Mvh, Dazl, Stra8, and Scp3).
Conclusion: It was found that although ADMSCs were attained easier and also cultured and differentiated rapidly, germ cell markers were expressed in BMMSCs significantly more than ADMSCs.

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