Anatomical Sciences Journal
Year:2007 | Volume:4 | Issue:17|Papers Count:5

Purpose: The aim of this study is to evaluate the effects of prepared serum from rat peripheral blood on growth and viability of rat bone marrow-derived mesenchymal stem cells (MSCs) in vitro.Materials and Methods: Bone marrow cells prepared from rat’s tibia and femur central canal were cultured either in medium containing fetal bovin serum (FBS) or rat peripheral blood-derived serum (PBDS) for three successive passages during which viability, growth and proliferation of the cells were evaluated by MTT [3- (4, 5- dimethylthiazol-2-yl)-2, 5- diphernyltetrazolium bromide], colony forming assay, calculation of population doubling number and daily examination of the cell growth for drawing growth curve. Each experiment was performed ten times and the mean values were statistically compared. In this study, third passaged cells were evaluated in terms of their bone and adipocyte differentiation potentials.Results: Morphologically, the cells cultured with PBDS appeared to be somewhat more spindle than of FBS cells. Comparatively, these cells seemed to have clear cut border than of FBS-cultured cells. According to MTT assay, the cells from PBDS group significantly showed more absorption value than of FBS group indicating that they would be more viable than FBS cells. Furthermore, in this group, more colon were formed and more population doubling number (about twice of FBS cells) occurred (p<0.004). Growth curve of two cells revealed further differences; in contrast to FBS, the cell from PBDS group showed no lag phase and their log phase was much longer than of FBS cells (2 versus 6 days). Third Passaged cells were readily differentiated to bone and adipocyte lineages confirming their mesenchymal stem cell nature.Conclusion: Taken together, the serum prepared from rat peripheral blood could significantly improve the viability and proliferation potential of the rat MSCs and in this term they could be considered as appropriate substitute for FBS.

Purpose: Parthenogenetic activation is among the crucial steps determining successful development of mammalian cloned embryos. This study, therefore, was conducted to evaluate the efficiency of a novel combined electrical-chemical artificial activation method for bovine cloning.Materials and Methods: In vitro matured bovine oocytes than were initially exposed to electrical pulse, used for cell fusion during cloning, and then treated with temporal sequential combinations of 3 chemical activators [calcium ionophore (CI), strontium (SR) and ethanol (ET)], followed by exposure to a protein kinase inhibitor or used for in vitro fertilization as control group. Treated and naturally fertilized oocytes were further cultured for up to 8 days. Embryo development was scored daily and blastocyst cell counting carried out using differential staining at day 8 of culture.Results: Among 15 temporal sequential combinations of three chemical activators, the best cleavage rates were associated with double (SR CI, 84.4%), triple (CI SR ET, 79.4%) and single (CI, 73.7%) compounds, respectively, which were not significantly different with each other and with in vitro fertilized (IVF) (85.5%). The highest blastocyst rates were gained with ET SR (25%), SR CI ET (21.7%) and CI (20.3%) accordingly which were not significantly different with each other but significantly lower than IVF (46%).Embryo cell counting further confirmed reasonably better quality of blastocysts produced using double, triple and single compounds.Conclusion: Although most of the sequential artificial activation compounds induced high cleavage rate close to IVF, but this did not assure comparable further embryo development to the blastocyst stage.Nevertheless, the results suggest exposure of in vitro matured bovine oocytes to electrical pulse, followed by exposure to CI 6- dimethylaminopurine (6-DMAP) or ET SR 6-DMAP could be regarded as the optimal artificial activation.

Purpose: This research has been carried out to study the morphological and ultrastructural changes of rabbit heart ganglionic neurons and nerve fibers after treatment with Amiodarone.Materials and Methods: 21 rabbits were divided into control and two experimental groups. The first experimental group for one week and the second experimental group for two weeks were injected intraperitoneally 80 mg/kg Amiodarone. The control group received normal saline the same dose as Amiodarone .Then the animals were anesthetized and perfused with Karnovsky, s fixative and the tissues were fixed and prepared for electron microscope study.Results: Ultrastructural changes in neurons and nerve fibers of experimental groups in comparison with control group included shrinkage of nuclear envelope and formation of electron dense masses in myelin of nerve fibers.Conclusion: These results indicate that Amiodarone causes ultrastructural changes in neurons and nerve fibers of intrinsic cardiac ganglia.

Purpose: To study the effect of ethanol on resumption of meiosis, maturation and development of immature mouse oocytes in vitro.Materials and Methods: Immature oocytes were dissected from ovary of NMRI strain mouse (6-8 weeks old) and divided into control and experimental groups. The absolute ethanol was imposed to oocytes at doses of 0 (Control), 0.05, 0.1, 0.2 and 0.4% (v/v) in maturation culture medium (MEM- + 100 mlIU/ml rFSH + 7.5 IU/ml hCG + 5% FCS). After 24 hours the matured oocytes were fertilized with spermatozoa in T6 medium and cultured for 5 days. Maturation and embryo formation observed by invert microscope and the results were considered as categorical variables with Chi- Square tests.Results: The percentage of resumption of meiosis and Metaphase II (MII) in control and the four experimental groups were 76.18%, 84.81%, 83.10%, 74.54%, 78.93% and 60.98%, 71.79%, 68.17%, 60.08%, 64.52%, respectively The percentage of the oocytes remained at the Germinal Vesicle (GV) stage were higher in 0.2 % (v/v) ethanol than the other groups (23.8%, 15.1%, 16.8%, 25.4%, 21%). The rate of resumption of meiosis and maturation mouse oocytes between control and experimental groups in 0.05 % (v/v) ethanol (P<0.001) and 0.1 % (v/v) ethanol (P<0.05) were significantly different. At 2-cell stage the rate of embryo formation between control and experimental groups in 0.2 and 0.4 %( v/v) ethanol were significantly different (P<0.001).Conclusions: The present study indicates that 0.05% (v/v) ethanol as solvent during in vitro culture media has positive effects on in vitro maturation of immature mouse oocytes, but same effects on embryonic development has not been detected. In conclusion it has not an inhibitory effect on embryonic development.

The median nerve is formed by fusion of lateral (C5-C7) and medial (C8, T1) cords from the brachial plexus. Bilateral variations in the formation of median nerve are very rare. One of such variations was observed during the dissection of the upper limb of approximately a 30-35-year-old man cadaver, according to Grant s method. The median nerve was formed by the union of three roots on both sides. There was an additional lateral root on both sides, while the first lateral root was short.On the left side, the second lateral root was short and united with the main trunk of median nerve proximal to the insertion of coracobrachialis muscle to humerus bone. However, on the right side, the second lateral root was long and pierced the coracobrachialis muscle accompanying with musculocutaneous nerve. Then it joined to the main trunk of the median nerve distal to the insertion of coracobrachialis muscle to humerus bone.Since, topographical situation of brachial plexus, median nerve and its communicating branches with musculocutaneous nerve have a very important role in evaluation of their injuries and neurological signs, awareness of the normal and variant patterns of the brachial plexus branches might be of great importance for medical specialists particularly anatomists and surgeons.