Anatomical Sciences Journal
Year:2006 | Volume:4 | Issue:2|Papers Count:10

Purpose: The purpose of this study was to isolate and purify fibroblastic cells from NMRI mouse bone marrow and evaluate their potential in differentiating among condrocytic cell lineage in vitro.Materials and Methods: The cells from NMRI bone marrow were harvested and plated at about 500 cells per well of 6-well plates for several days. The pure fibroblastic cells were not appeared until the second passages of the cells were performed. To differentiate into cartilage, two different techniques were employed. In micro mass culture technique, about 200000 fibroblastic cells pletted by centrifuging were incubated in chondrogenic medium for three weeks. In Monolayer Culture technique, about 200000 cells seeded in 24-Well plate were provided with chodrogenic medium for three weeks. In the end of differentiation period, the cells were evaluated by histochemistry and RT-PCR analysis for cartilage differentiation. To examine the mesenchymal nature, the isolated cells were differentiated into bone and adipocytic cell lineages.Results: In the early days of primary culture, the dominant population of the cells was that with spindle shape morphology that expanded in subsequent subculture and almost purified in second passage.Differentiation results showed that in contrast to monolayer culture system in which chondrogenic differentiation was not occurred in micro mass system the cells were apparently differentiated into cartilage as it was evidenced by our evaluation. RT-PCR analysis was indicative of high production of collagen type II, X and aggreacan among the differentiated cells and histochemistry results showed the methachromatic matrix accumulation between the cells. The isolated cells in this study were mesenchymal cells as they readily differentiated into bone and adipocyte.Conclusion: The cells isolated by low-density culture system could be differentiated among the chondroblastic lineage and have mesenchymal characteristic.

Purpose: The present study intends to show the characteristics of cultured limbal stem cell (CLSCs) and to compare them with normal Conjunctival (C), Limbal (L) and Cornea (K) tissues.Materials and Methods: The expressions of a set of genes potentially involved in differentiation and stemness function of limbal stem cells were assessed in freshly prepared limbal, corneal, and conjunctival tissues by Immunocytochemistry, Immunoblot and RT-PCR. Also, ultra structure of limbal explants was cultured on human amniotic membrane (HAM) and normal tissues were evaluated by scanning electron microscopy and transmission electron microscopy.Results: PAX6 and OCT4 were expressed in limbal, conjunctival, and corneal tissues. However, K3 were expressed only in corneal and K12 in both tissues. The gene, K3, was not expressed in limbal cultured explants (LCEs), whilst OCT4 was expressed in all stages. The expression of K12 and Cx43 increased over time in-vitro. Whereas, p63 expression was reduced. Ultrastructure analysis of the LCEs showed typically immature organization of intracellular organelles and architecture.Conclusions: Our data suggest that limbal tissues are heterogeneous; containing both precursor cells and some differentiated epithelial cells. An epithelial cell sheet grown from a limbal explant on HAM retains important physiological and structural characteristics almost similar to normal limbal epithelium. Although specific markers of LSCs still remain to be identified, there is an increasing need to also determine factor(s) involved in their stemness.

Purpose: This work studies possible role of FOS proteins, retinoic acid (RA) and mitogens during optic vesicle development in vitro.Materials and Methods: Fos was detected as a product of 55-62 kDa in optic vesicles via electrophoretic procedures. Incubation (24hr) of optic vesicles with RA 25nM and mitogens(serum10%,insulin2.5%) on the day 9.5 of gestation revealed that Fos was induced by mitogens like serum and insulin.Results: The results suggested that the regulation of fos may be required for cell proliferation and differentiation in normal eye development. Fos induction was inhibited by retinoic acid.Conclusion: the results showed that retinoic acid prevents vesicles growth and reduces rate of protein significantly in all treatment groups and that in addition to Fos mitogens are essential to growth of vesicles.

Purpose: the aim of this study is to analyze the factors contributing to the success of frozen- thawed embryo transfer (ET).Material and method: This retrospective, case-control study was carried out on 700 infertile couples treated with IVF and ICSI and had embryo freezing in Isfahan Fertility and Infertility Center, Iran, from the beginning of July 2000 to the end of June 2003.Results: factors affecting the success rate of frozen- thawed ET are: the number of 7-8 cell embryos with survival rate > 75%, the ratio of frozen-thawed embryos with survival rate > 75% to the total number of frozen-thawed embryos, the number of frozen embryos, mean percentage viability ,cleavage stage of fresh embryos, and mean viability percentage of frozen- thawed embryos. Significantly correlated factors with the number of frozen-thawed 7-8 cells embryos with survival rate > 75% are the number of type A fresh 7- 8 cell embryos, fresh embryo survival rate and cellular staging, the number of embryos on the third day following insemination, fresh embryo survival coefficient, the number of retrieved oocytes, and maternal age (with a negative correlation).Discussion: increasing the viability percentage and number of cryopreserved embryos increases the success of frozen-thawed ET. Among all, the number of 7-8 cell embryos with survival rate > 75% are the most influential ones. Also, improving the before freezing such as the number of retrieved oocytes and the number and quality of embryos improves the viability percentage and number of frozen- thawed embryos.

Purpose: Present study was designed to evaluate the potential of co-culture systems to overcome deleterious effect of exposure of mouse 2-cell embryos to low temperature.Materials and Methods: 2-cell embryos were flushed from oviduct of super ovulated NMRI mice into HTF medium with 15% BSA. After washing 3 times with HBSS and with 15% BSA, embryos were exposed to laboratory temperature (LT) (22-24 ° C) for 1, 3 and 5 hours. Embryos in each group were simultaneously transferred into drops of HTF, MEM-a as control as well as Mouse Embryonic Fibroblast (MEF) and Human Embryonic Fibroblast (HEF) as treatment. All the embryos were incubated in humidified 37° C incubator with 5% CO2 in air for 120 hours.Experiments were replicated 6 times and development of embryos was recorded every 24 hours for 5 days. Data were analyzed with c2 test and any statistic difference with p<0.05 was considered significant.Results: Development of the embryos following 5 hour exposure to LT decreased significantly (p<0.05) after 4 and 5 days cultivation in HTF and MEM-a when compared to control. Co-culture of the embryos with either MEF or HEF increased the rate of blastocyst formation in all the groups that had been exposed to LT. No significant difference was noted between the embryos in co-cultured control group and experimental groups. A sharp increase in development of the embryos, which were exposed to LT for 5 hours and co-cultured with the feeder cells after 4 and 5 days, was observed when compared with MEM- a group (p-<0.05).Conclusions: We conclude that mouse 2-cell embryos can withstand deleterious effect of LT for less than 3 hours. However, keeping embryos for 5 hours at LT decreases the rate of development. Either cell employed in present study enhanced development of embryos especially when the embryos were exposed to LT.

Purpose: The purpose of this study was to investigate the expression of Bax and surviving after cold injury and the ability of magnesium chloride to inhibit of apoptosis. Materials and Methods: To produce cold injury, a metal probe cooled with liquid nitrogen was applied to the surface of the mouse intact skull above the parietal lobe by force of 100 gr for 30 sec. Brains were removed 72 h after cold injury and sections were prepared to study immunohistochemistry reactions of Bax and Survivin antibodies. The data were analyzed using t- tests.Results: The results indicated that expression of Bax was higher in model group comparing with treated ones. Survivin was expressed more in the treated group and there was significant difference with model group (p<0.05).Conclusion: MgCL2 with optimum dose causes activate expression of Survivin and inhibition of apoptosis.

Purpose: Hydrogen peroxide is an unstable, colorless, heavy liquid used as bleach in industry and as an antiseptic in house holds. Teaching human anatomy, at any level, relies not only on the expertise of a tutor but also on the availability and use of good teaching aids. Plastination specimens have unique position as teaching aids to exhibit accurate anatomical structures and they are also easy to store and handle by students. Darkening of plastinated tissue is one of the problems that may occurr because dark tissue is unusable. We applied this survey to examine clearing of the hydrogen peroxide on posterior wall of trunk, spinal cord and medulla oblongata before Plastination.Materials and Methods: This study was carried out on a human body after fixation, dissection, dehydration and defatization of the cadaver. The hydrogen peroxide was prepared in several concentration (33.3%,16.6%,8.3%,4.1%) was added to the specimen of master specimen at several times (1 ,2, 4,6,8,10,12,18 and 24 hours) for clarification. We choused (16.6% per 6hour) as the best situation for clarify of specimen. After clarification of original specimen, impregnation and curing was carried out. Finally, the prepared Plastinated specimen was compared with the standard Plastinated model.Results: The obtained Plastinated specimen was compared with the standard model of Heidelberg Germany for appearance and resolution. The plastinated specimen obtained was compared with the standard model of Heidelberg, Germany for its stature, flexibility and traction by the universal test DARTC (England) apparatus in the Department of Medical Physics, Isfahan School of Medicine. The result obtained was found to show that the P-value was. 08 and the standard error was (-0.460±0.382).Conclusion: The Plastinated specimen prepared, provides an excellent opportunity to demonstrate and study the dissected areas of the difficult structures which can be of great benefit in teaching gross anatomy and neuro-anatomy, In addition, because of the durability, safety, reduction intoxic and noxious fumes of formalin, the Plastinated specimens can be unique materials as a teaching aid by the side of (Wet) specimens.

Since "variation" is one of anatomy topics which helps other medical sciences, specially surgery, and gives assures surgeons of surgical operations. Thus, investigation of variations in cadavers is very important and essential.A cadaver about 50-year-old and white race was prepared and dissected at Anatomy Department of Fasa Medical School.After dissecting the left arm, it was revealed that in the half of the arm brachial artery has been divided into its terminal branches which are radial and ulnar arteries. The radial artery was located in the medial and the ulnar artery was located in the lateral side so that ulnar artery supply blood to biceps muscle. Coracobrachialis muscles are also supplied by radial artery. Also, brachialis muscles are supplied by ulnar and radial arteries respectively. The profunda brachei was laterally branched immediately over the point of dividing of the brachial artery. The radial and ulnar arteries were crossed in about 3cm over the top of cubital fossa. Radial artery extended toward extensors and ulnar artery extended toward flexors groups muscles .These variations can make problems for surgeons in arm surgery.