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Article
Flowcytometry for Estimating Efficient Transfection of Bone Marrow Cells with BCR-ABL Gene

Authors: Rihab Nasr --- Manal Adnan Habib --- Ahmad Iskandarani
Journal: Iraqi Academic Scientific Journal المجلة العراقية للاختصاصات الطبية ISSN: 16088360 Year: 2012 Volume: 11 Issue: supplement Pages: 649-653
Publisher: The Iraqi Borad for Medical Specialization المجلس العراقي للاختصاصات الطبية

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Abstract

ABSTRACT:BACKGROUND:Chronic myeloid leukaemia can be accurately modeled in laboratory mice by the retroviral transfer of a BCR-ABL gene into murine hematopoietic stem and progenitor cells, followed by transplantation of these cells into irradiated recipient mice.OBJECTIVE:The use of Retroviral vector for transfer of BCR-ABL gene into murine bone marrow cells (BMC) and measurement of efficiency of transfection by flowcytometry.METHODS:Murine bone marrow cells obtained from mice were cultured in a medium containing the supernatant of BCR-ABL (p210) transfected platinum E cells which is rich in retroviral vector carrying the BCR-ABL (p210) gene.The vector express green fluorescent protein (GFP) as well so that the efficiency of transfection of murine BMC with the target gene was able to be measured using flowcytometry.RESULTS:The use of the retrovirus packaging cell line enhanced the transduction of BMC with the retroviral vector and efficiency of transfection was 72% as measured by the flowcytometry.CONCLUSION:Transfer of BCR-ABL gene into murine BMC by retroviral vector that carry GFP marker which allowed the estimation of transfection efficiency by the flowcytomery


Article
Isolation and characterization of bone marrowmesenchymal stem cells from rat and rabbit; a modified method
عزل وتوصيف خلايا نخاع العظم ألجذعيه الميزنكيميه من الجرذانوالأرانب .طريقة محورة

Authors: Limes HusamAlmanseekanaa --- Ali MansoorJasim --- RaedHamzah Mohammed
Journal: karbala journal of pharmaceutical sciences مجلة كربلاء للعلوم الصيدلانية ISSN: 70272221 Year: 2012 Issue: 3 Pages: 44-51
Publisher: Kerbala University جامعة كربلاء

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Abstract

Bone marrow MSCs were obtained from New Zealand White rabbits (~3 kg, male) and also from white rat (Ratusratus) by the Guideline for Animal Care and use Committee for Teaching and Research using a modified procedure (1,2) Briefly, after the animals was sacrified with carbon monoxide cabinet, tibias and femurs were excised and the bone marrow was extracted under sterile conditions.The collected marrow was mixed and dispersed with PBS. After that the (MSCs) were separated by density-gradient centrifugation over ficoll. Viability of the separated MSCs was determined via trypan blue staining technique. Then, cells were seeded into ten tissue culture flasks containing complete culture medium suplimented with 10% fetal bovine serum (FBS) and antibiotics, incubated at 370C in a humidified atmosphere with 5% CO2. Once the colonies reached 80–90% confluence, they were ready to be detached with trypsin/EDTA and suspended in medium for continuous culture

تضمنت الدراسة عزل و تنمية الخلايا الجذعية الميزنكيمية من نقي العظم للارنب الابيض الذكر والذي يزن حوالي 3 كغم والجرذ الابيض المختبريين طبقا لقوانين هيئة العناية بالحيوان واستعماله للأغراض البحثية و التعليمية وذلك باتباع طريقة محورة لهذا الغرض. بأيجاز, بعد قتل الحيوان داخل كابينة غاز أحادي أوكسيد الكاربون تم استئصال عظمي الفخذ وعظمي القصبة وبعدها تم استخراج نقي العظم منها والذي خلط مع محلول الفوسفات الملحي ( (PBS. ثم تم فصل الخلايا الجذعية الميزنكيمية من الخليط بوساطة السنترفيوج مع فارق الكثافة وذلك بأضافة مادة الفيكول. بعد ذلك تم تحديد حيوية الخلايا المفصولة عن طريق تقنية التصبيغ بصبغة التريبان الزرقاء. بعدها تم استنبات الخلايا في عشرة اطباق زرعية نسيجية يحوي كل منها على الوسط الزرعي النسيجي المتكامل مدعما بالمصل البقري والمضادات الحيوية حيث تم الحضن بدرجة 37 مئوية في محيط رطب يحوي 5% غاز ثاني أوكسيد الكاربون. عندما أصبحت درجة اتصال المستعمرات الخلوية 80-90% كانت جاهزة لفصلها بواسطة محلول أنزيم التربسين مع EDTA واستنباتها في الوسط الزرعي الدائمي


Article
Facilitating osteogenesis of Hydroxyapatite granules by Autogenous bone marrow

Author: Dr .Hareth.H.Kaskos, MS.c oral and maxillofacial surgery, Lecturer د. حيدر
Journal: MUSTANSIRIA DENTAL JOURNAL مجلة المستنصرية لطب الاسنان ISSN: 18138500 Year: 2012 Volume: 9 Issue: 1 Pages: 83-88
Publisher: Al-Mustansyriah University الجامعة المستنصرية

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Abstract

The aim study to evaluate whether Hydroxyapatite granules alone or in combinationwith autogenous bone marrow can facilitate bone formation. Materials and Methods: (8)dogs of both sexes were used as a model in this experimental study ,they were divided intotwo groups , the control group in which ( 4) dogs were operated on in this group receivedHydroxyapatite granules alone in defect created in the body of mandible (1cm) in diameterand experimental group in which (4)dogs were operated on in this group receivedHydroxyapatite granules mixed with autogenous bone marrow implanted in the defect alsocreated in the body of mandible (1cm) in diameter .Alkaline phosphatase enzyme (ALP)used as parameter to evaluated the degree of osteogenesis ,so blood sampling from femurvein was aspirated at 1st ,2nd ,3rd and 4th weeks respectivelly for biochemical analysis.Results :Statistical analysis showed that there is significant elevation of serum (ALP) inexperimental group at day 15 and 21 comparing to control group .Conclusion:Hydroxyapatite with autogenous bone marrow than it is used alone. has great effect onfacilitating of osteogenesis.


Article
Evaluation the effect of autologous bone marrow – derived mesenchymal stem cells as a treatment in diabetic rabbits

Authors: Mohamed Abdul-Hameed Mohamed محمد عبد الحميد محمد --- Wasan H. Younis وسن يونس --- Nahi Y.Yaseen ناهي ياسين
Journal: Journal of baghdad college of dentistry مجلة كلية طب الاسنان بغداد ISSN: 16800087 Year: 2012 Volume: 24 Issue: special issue 2 Pages: 55-60
Publisher: Baghdad University جامعة بغداد

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Abstract

Back ground: Type 1 diabetes is the result of an autoimmune attack against the insulin-producing beta cells of thepancreas. Current treatment for patients with type 1 diabetes typically involves a rigorous and invasive regimen oftesting blood glucose levels many times a day along with injections of recombinant insulin. Many recent researcheshave shown that stem cell therapy can be the best choice for treatment of this disease. The aims of this researchwere investigating regeneration of pancreatic beta cells of type 1 diabetic rabbits after stem cell transplantation.Materials and Methods: 32 rabbits weighting an average of (2.5 - 3 kg) were used in this experimental study, anddivided into 2 groups as follows; group A ( contains 16 controlled diabetic rabbits received insulin as a treatment )and group B ( contains 16 diabetic rabbits received autologous mesenchymal stem cells as a treatment).Theinduction of diabetes was achieved by a single dose of intravenous injection of the Alloxan, which was administeredto the rabbits via the marginal ear vein, mesenchymal stem cells were differentiated into insulin – producing cells andreimplanted into the rabbits of group B with daily monitoring of blood glucose level and body weight.Results: The insulin – producing cells regulated the hyperglycemia resulted from diabetic rabbits , 7 to 9 days afterreimplantation the blood glucose level were decreased from about( 400 mg/dl into 180 mg/dl).Conclusions: Islet-like functional cells can be differentiated from bone-marrow mesenchymal stem cells (MSCs),which may be a new procedure for clinical diabetes stem -cell therapy, these cells controlled blood glucose level indiabetic rabbits as the effect of insulin. MSCs play an important role in diabetes therapy by islet differentiation andtransplantation.


Article
Effect of auto-transplantation of bone marrow on the nerve autography in the dogs
تأثير الزرع الذاتي لنخاع العظم على الترقيع الذاتي للعصب في الكلاب

Author: O.H. Al-Hyani أسامة حازم الحياني
Journal: Al-Qadisiyah Journal of Veterinary Medicine Sciences مجلة القادسية لعلوم الطب البيطري ISSN: 18185746 23134429 Year: 2012 Volume: 11 Issue: 2 Pages: 128-141
Publisher: Al-Qadisiyah University جامعة القادسية

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Abstract

The research was conducted to study the repair of transected sciatic nerve through by grafting sciatic nerve with nerve segment and addition of bone marrow in dogs. Twelve adult dogs from both sexes was used. They was divided into two groups, six animals in each group. The left sciatic nerve was used as a model in this research for monitoring the process of nerve repair. In group one, the sciatic nerve was transected and a piece of nerve about 1cm in length was removed, and the resultant gap was repaired by autotransplantation with a segment of nerve harvested from the median nerve of the forelimb with the consideration that the harvested median nerve segment was slightly longer than the resected sciatic nerve segment. The implanted nerve segment was sutured with sciatic nerve using non absorbable suture (nylon 5). In group two, the same surgical procedure was performed as in group one, but a bone marrow that aspirated from the same animal was applied on the nerve transplantation. The assessment of sciatic nerve repair was accomplished by studying the clinical observation of normal physiological function of the operated limb, additionally studying the histological changes on the nerve graft transplantation at 30 and 45 postoperative days. The study was revealed, the application of bone marrow on the nerve graft segment was enhanced the degree of healing of transected sciatic nerve that indicated by improvement the functional use of affect hind limb clinically, with improvement the vasculrization of nerve graft segment and increase proliferation of nerve cells (Schwann and microglial cells) with extension of collagen fibers that aid to bridge the sciatic nerve with grafted nerve segment histologicaly rather than in group one.In conclusion the addition of bone marrow on the nerve graft segment accelerate the degree of healing of transected sciatic nerve with improve the functional use of operated limb.

صمم هذا البحث لدراسة إصلاح العصب ألوركي المقطوع من خلال ترقيع العصب ألوركي بقطعة من العصب مع إضافة نخاع العظم في الكلاب. اثنا عشر كلبا ومن مختلف الجنسين استخدمت في هذا البحث,حيث قسمت الحيوانات إلى مجموعتين وبواقع ست حيوانات لكل مجموعة. تم اختيار العصب ألوركي الأيسر كنموذج لمعرفة سير خطوات الالتئام في العصب. ففي المجموعة الأولى تم قطع العصب ألوركي وإزالة قطعة منه بطول 1 سم. بعدها تم إصلاح الفسحة الناتجة في العصب ألوركي من خلال الزرع الذاتي لقطعة عصب تم أخذها من العصب الإنسي للقائمة الأمامية وبطول أكثر قليلا من طول القطعة التي أزيلت من العصب ألوركي. تم خياطة قطعة العصب المزروعة مع العصب ألوركي باستخدام خيط غير ممتص (نايلون 5). إما في المجموعة الثانية فتم استخدام نفس الخطوات الجراحية التي استخدمت في المجموعة الأولى مع إضافة مادة النخاع العظمي المأخوذة من نفس الحيوان على قطعة العصب المزروعة. تم تقيم حالة التئام العصب ألوركي من خلال دراسة الوظيفة الفسلجية الطبيعية للطرف المصابة سريريا مع دراسة التغيرات النسيجية الحاصلة في قطعة العصب المزروعة كل 30و45 يوم. أظهرت الدراسة أن إضافة نخاع العظم على العصب المزروع حسن من التئام العصب ألوركي المقطوع من خلال تحسن الاستخدام الوظيفي للطرف المصابة سريريا مع تحسن التجهيز الدموي للزرعة وزيادة تكاثر الخلايا العصبية (الخلايا الدبقية الدقيقة وخلايا شوان ) وامتداد الألياف الغراوية التي ساعدت على ارتباط قطعة العصب المزروعة مع العصب ألوركي نسيجيا على عكس المجموعة الأولى .بالاستنتاج نجد إن إضافة نخاع العظم على قطعة العصب المزروعة يحسن من التئام العصب ألوركي المقطوع مع تحسن الاستخدام ألسريري للطرف المصابة.

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