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Article
STUDY THE GENOTOXICITY EFFECT OF HOECHST 33258 STAIN IN THE HUMAN LYMPHOCYTE
دراسة التأثير السمي الوراثي لصبغة هوكست Hoechst 33258 في الخلايا

Author: Muhammad A. Al-A’adhmi
Journal: Iraqi Journal of Biotechnology المجلة العراقية للتقانات الحياتية ISSN: 18154794 Year: 2012 Volume: 11 Issue: 2 Pages: 209-218
Publisher: Baghdad University جامعة بغداد

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Abstract

Most stains using in DNA labeling has the ability to cause genetic mutations because of its ability to intercalated on DNA. Hoechst 33258 stain is one of a widely used stains in laboratory preparations, some studies indicated that this stain caused genetic mutations. It is important to detect the relationship between this stain and the people who works in laboratories, there is a lack of studies describing accurately the genotoxicity of this stain. This study is an investigation on the genotoxicity of Hoechst 33258 stain in human lymphocytes (in vitro) using cytogenetic tests including Mitotic Index (MI), Chromosome Aberration (CA), Sister chromatid exchange (SCE) and Preparation of micronuclei, using different concentration of Hoechst stain 5μg/ml, 10μg/ml, 15μg/ml, 20μg/ml. The results showed significant increase in Mitotic Index (MI), Chromosomal Aberration (CA), Sister chromatid exchange (SCE) and Micronuclei this increase is direct proportion with the increase of stain concentration. The Chromosome Aberration represented by chromosomal and chromatid breaks showed at (5,10,15,20)μg/ml, chromosomal and chromatid gap showed at (5,10,15,20) μg /ml, while ring chromosome and dicentric chromosome showed at (15,20) μg /ml.

من المعروف إن معظم الصبغات المستخدمة في وسم المادة الوراثية (DNA) تكون ذات قابلية على إحداث الطفرات الوراثية لما تمتلكه هذه المواد من صفات تمكنها من الارتباط بالـ(DNA) ومن بين تلك الصبغات هي صبغة هوكست Hoechst 33258 والتي لها إستعمالات مختلفة في التحضيرات المختبرية الخاصة بالمادة الوراثية، تشير بعض المصادر على إمتلاكها القابلية على إحداث الطفرات الوراثية، ولأهمية الموضوع وعلاقته بتماس مباشر مع العاملين في المختبرات وإستعمالهم لهذه الصبغة ولعدم توفر دراسات تصف بشكل دقيق التأثير السمي الوراثي لهذه الصبغة جاءت أهمية هذه الدراسة، حيث هدفت الى الكشف عن التأثير السمي الوراثي لصبغة الهوكست في الخلايا اللمفاوية للانسان (خارج الجسم الحي) وذلك بإستعمال فحوصات الوراثة الخلوية المشتملة على إختبار معامل الانقسام الخلوي وإختبار الزيغ الكروموسومي وإختبار التبادل الكروماتيدي الشقيقي وإختبار تكون الأنوية الصغيرة مستخدمين تراكيز مختلفة من صبغة الهوكست وهي على التوالي: (20, 15, 10, 5) مايكروغرام/مليلتر. إذ سجلت أنواع من التغيرات الكروموسومية منها كسر كروماتيدي وكسر كروموسومي في التراكيز (20,15,10,5) مايكروغرام/مليلتر كما سجلت تغيرات من نوع فجوة كروماتيدية وفجوة كروموسومية في التراكيز (0،20،15،10،5) مايكروغرام/مليلتر، أما التغيرات الكروموسومية من نوع قطع كروموسومية بدون مرتكز وكروموسوم حلقي كروموسوم ثنائي المرتكز سجلت في العينات المعاملة بالتراكيز (15و20) مايكروغرام/مليلتر.


Article
Effects of Exhaustive Exercise on Peripheral Blood Lymphocytes

Authors: Israa F. Ja'afar اسراء فائق --- Balsam F. Abid بلسم فاروق
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: PISSN: 00419419 / EISSN: 24108057 Year: 2010 Volume: 52 Issue: 1 Pages: 84-88
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Acute exercise stress is associated with a lower lymphocyte functional response. It is suggested that exercise can enhance or reduce immunity depending on the frequency, duration, and intensity of the exercise.Subjects and Methods: From November 2007 to January 2008, thirty healthy Volunteers were recruited from medical, paramedical staffs, and students of Baghdad College of Medicine (all males). Participants were chosen according to the following: (1)Age (between 20 and 35 years), (2) Non obese : Body mass index (BMI in kg/m2) was used as an index of obesity, (3) Non-smoker, (4) No recent medical problem, (5) No recent medications, and (6) No musculoskeletal complaint. After a general medical checkup, the thirty subjects were subjected to the exercise protocol. Blood samples were taken before, and immediately after the end of exercise. Results: The mean age of individuals enrolled in the study was 29 ± 4years. Exercise induced significant granulocytosis whereas lymphocytes declined to a level significantly below the baseline. Trypan blue exclusion test showed that the initial percentage of nonviable lymphocytes was 2.90% ± 0.2%. After exhaustive exercise, cell viability decreased where percentage of nonviable cells was 4.3% ± 0.3% (p<0.005). Early apoptotic lymphocytes showed membrane blebbing and did not take the dye, while dead cells appeared blue from dye uptake. MDA was 3.0 ± 0.23 nmol/ml. An increment in the mean value of MDA was observed in the post exercise blood sample that was withdrawn immediately after exercise (p<0.005).Conclusion: The present study shows that exhaustive exercise induces apoptosis in peripheral blood lymphocytes manifested by increased level of malondialdehyde, an oxidative stress marker. This finding supports the hypothesis that apoptosis could be involved in exercise-induced lymphocytopenia and this effect depends on exercise intensity. Keywords: Apoptosis, Exercise, MDA, lymphocyte.

Keywords

Apoptosis --- Exercise --- MDA --- lymphocyte.


Article
Effect of Moderate Exercise on the Level of Melatonin Hormone and Lymphocyte Apoptosis in Healthy Subjects

Authors: Israa F. Ja'afar --- Huda Arif Jassim --- Alaa G. Hussein
Journal: Iraqi Academic Scientific Journal المجلة العراقية للاختصاصات الطبية ISSN: 16088360 Year: 2010 Volume: 9 Issue: 1 Pages: 88-94
Publisher: The Iraqi Borad for Medical Specialization المجلس العراقي للاختصاصات الطبية

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Abstract

ABSTRACT: BACKGROUND: Physical exercise is important for promotion of the immune system. The immune responses to exercise are dependent on the intensity and duration of the exercise performed. During exercise, metabolic and hormonal changes occurred that can induce lymphocyte apoptosis. Melatonin is one of the hormones that are affected by exercise, it is a pineal gland hormone, its production and secretion followed a circadian pattern, onset of darkness stimulate its secretion, while day light suppress its production. Melatonin is regarded as an anti-aging hormone; it has a role in the elimination of harmful lymphocytes which may give rise to chronic inflammation and possibly to autoimmunity. OBJECTIVE: This prospective study aimed to elucidate the effect of moderate endurance exercise on melatonin hormone and the percentage of peripheral blood lymphocyte apoptosis in healthy subjects. METHODS: Sixty healthy subjects with a mean age of (37.05 ± 13.02) year, participated in the study. They were subjected to moderate exercise session which lasted for 60 min and repeated 3 times per week for 3 months. The exercise intensity was 50-60% of heart rate reserve (according to Karvonen formula). Two exercise stages were performed; the first stage include 30 minute of physical activity program and the second stage include 30 minute of treadmill exercise. Blood sample were collected before and after exercise to examine its effect on the level of melatonin hormone and apoptosis of peripheral blood lymphocyte. RESULTS: This study showed significant increment in the level of melatonin hormone and the percentage of lymphocyte apoptosis after exercise in comparison to pre exercise values for each week along the period of the test (P < 0.05). Three months exercise showed a significant correlation with the levels of melatonin hormone (P= 0.008, r= 0.44), and with percentage of lymphocyte apoptosis (P = 0.001, r= 0.701). CONCLUSION: It can be concluded that moderate exercise for three months was associated with increased percentage of periphral blood lymphocyte apoptosis and increased level of melatonin hormone in the first two months while in the third month this anti aging hormone had reached a steady state after the subjects get trained to exercise


Article
The effect of Serratia marcescens protease on human lymphocytes transformation
تأثير إنزيم البروتييز لبكتريا ال Serratia marcescens على تحول الخلايا اللمفاوية لدم الإنسان

Authors: Raghd H.Mahdi رغد حربي مهدي --- Shahla A. Hasan شهلاء علي حسن --- Jinan M. Hasan جنان محمد حسن
Journal: Iraqi Journal of Science المجلة العراقية للعلوم ISSN: 00672904/23121637 Year: 2013 Volume: 54 Issue: 3 Pages: 531-535
Publisher: Baghdad University جامعة بغداد

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Abstract

Fifteen blood samples were collected from healthy males and females (6 males & 9 females), average age (21-34 years) in heparinized sterile tubes. The extracellular protease was extracted from a clinical isolate of Serratia marcescens that was isolated from a patient suffering from urinary tract infection taken from the Central Health Laboratory. The extracted protease was purified partial by two steps, precipitation with 30-55% saturation of ammonium sulfate following with dialysis and ion exchange chromatography DEAE-cellulose. The protease concentration was 0.15 mg/ml. Two concentration 0.25µg/ml and 0.5µ/ml of protease were prepared and applied in current study. Lymphocyte transformation test using whole blood was used in order to study the effect of protease concentrations on human lymphocyte transformation. The results show that these were highly significant differences (P < 0.05) between control samples and lymphocytes stimulating with Phytohemagglutinin (PHA) and between control sample and 0.5µg/ml protease,; while there was no significant differences (P < 0.05) between control sample and 0.25 µg/ml protease concentration . That is mean that the higher concentrations of protease have the same effect of PHA on human lymphocyte transformation and it is better in effect on human lymphocyte transformation than the lowest concentration

جمعت خمسة عشر عينة دم من ذكور و إناث أصحاء (6 ذكور و 9 إناث) معدل العمر يتراوح بين (21- 34 سنة) بواسطة أنابيب معقمة حاوية على مادة الهيبارين. تم استخلاص الانزيم الحال للبروتين (البروتييز) من عزلة مرضية لبكتريا ال Serratia marcescens من مريض يعاني من التهاب المجاري البولية من مختبر الصحة المركزي. و تم تنقيته بخطوتين تضمنت الترسيب باستخدام سلفات الامونيوم المشبعة بنسبة اشباع 30-55% و ديلزة و الترشيح باستعمال المبادل الأيونيеsolullеC -EAΕD . وقد كان تركيز البروتين 0٫15 ملغم/مل ̦ ومن هذا التركيز تم تحضير التراكيز (0٫25 مايكرو غرام/مل , 0٫5 مايكرو غرام/مل). استعمل اختبار تحول الخلايا اللمفاوية لدراسة تأثير تركيز الانزيمات الحالة للبروتين على تحول الخلايا اللمفاوية لدم الإنسان, و قد اوضحت النتائج ان الفروقات كانت جدا معنوية(P < 0.05) بين عينات السيطرة و الخلايا اللمفاوية المحفزة بالمشطر النباتي(AΗΡ), وبين عينات السيطرة و تركيز0,5 مايكروغرام/مل لانزيم البروتييز, و في حين لا توجد فروقات معنوية(P < 0.05) ما بين عينات السيطرة وتركيز0,25 مايكروغرام/مل من انظيم البروتييز مما يدل على ان التراكيز العالية لانزيم البروتييز لها فعل مشابه للمشطر النباتي(AΗP) على تحول الخلايا اللمفاوية لدم الإنسان كما ان لها تأثير أكبر من التراكيز الواطئة على تحول الخلايا اللمفاوية لدم الإنسان.


Article
Effect of Moderate Exercise on the level of DHEAS hormone and lymphocyte apoptosis in healthy subjects

Authors: Alaa G. Hussein علاء حيبن --- Israa F. Ja'ffar اسراء فائق --- Huda A. Jassim هدى جاسم
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: PISSN: 00419419 / EISSN: 24108057 Year: 2010 Volume: 52 Issue: 2 Pages: 225-229
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Physical exercise is essential for performance of the immune system. The integrity of immune response is dependent on the intensity and duration of the exercise performed. During exercise the metabolic and hormonal changes that occurred can induce lymphocyte apoptosis. Dehydroepiandrosteron sulphate (DHEAS) is one of the hormones that is affected by exercise, it is an androgen released from the adrenal cortex in response to adrenocortiocotropin releasing hormone (ACTH). DHEAS is regarded as an anti-aging hormone; it is primarily immune enhancing, providing some degree of counterbalance to the potentially detrimental effects of catecholamine and cortisol which are immunosuppressive if chronically elevated, it also had a role in the elimination of harmful lymphocytes which may give rise to chronic inflammation and possibly to autoimmunity. Methods: Sixty healthy subjects were involved in this study with a mean age of (37.05 ± 13.02) year. They were subjected to moderate exercise session which lasted for 60 min and repeated 3 times per week for 3 months. The exercise intensity was 50-60% of heart rate reserve (according to Karvonen formula). Two exercise stages were performed; the first stage include 30 minute of physical activity program and the second stage include 30 minute of treadmill exercise. Blood samples were collected before and after exercise to examine its effect on the level of DHEAS hormone and apoptosis of peripheral blood lymphocyte. Results: This study showed significant increment (P < 0.05) in the level of DHEAS hormone and the percentage of lymphocyte apoptosis after exercise in comparison to pre exercise values for each week along the period of the test.Three months exercise showed a significant correlation with the levels of DHEAS hormone (P= 0.000, r= 0.75), and with percentage of lymphocyte apoptosis (P = 0.001, r= 0.701).Conclusion: in this prospective study, following moderate exercise for three months; there was increase in the percentage of periphral blood lymphocyte apoptosis and increased level of DHEAS hormone in the first two months while in the third month this anti aging hormones had reached to a steady state after the subjects get trained to exercise. Keywords: moderate exercise, DHEAS, lymphocyte apoptosis.


Article
The role of Bcl-2 protein, Bax protein and there ratio in peripheral bloodlymphocytes of asthmatic patients

Authors: Haider S. Kadhim --- Musaab H Ali --- Israa F. Jaffar
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: PISSN: 00419419 / EISSN: 24108057 Year: 2008 Volume: 50 Issue: 2 Pages: 219-222
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Asthma is an inflammatory airway disease; this inflammatory response can
be attributed to reduced lymphocyte apoptosis in peripheral blood and in airway tissues.
The mechanism behind this could be attributed to decreased Bcl2 protein and increase
Bax protein in peripheral blood lymphocytes of asthmatic patients.
Aim: to explore the mechanism behind decreased lymphocyte apoptosis in peripheral
blood of asthmatic patients at cellular level.
Method: Ninety four subjects; (44) control and (50) patients were included in this study
during the period from (2003) to (2004). The aspirated lymphocytes for each individual
were prepared and stained by immunocytochemistry to study the percentage of anti-
apoptotic Bcl-2 protein, pro-apoptotic Bax protein and their ratio.
Results: Our results showed that the percentage of Bcl2 protein in peripheral blood
lymphocyte of asthmatic patients were significantly higher than control (33.23±11.56,
21.47±4.16)(P=<0.00001).
The percentage of Bax protein in peripheral blood lymphocyte in asthmatic patients was
significantly lower than the control (21.43±6.88, 24.89±4.8) (P-O.0323). Bcl2/Bax ratio
was positive in asthmatic patients there was significant difference in comparison with the
control (1.68±0.57, 0.87 ± 0.17) (P=<0.00001).
Conclusion: This study clarifies the role of these 2 proteins in the process of reduction of
apoptosis in peripheral blood lymphocytes of asthmatic patients.


Article
Sex Chromatin in New Hampshire Cocks
الصبغين الجنسي في سلالة النيوهمشاير في الديكة

Author: Maha K. Abd al-Kareem مها خالد عبد الكريم
Journal: Baghdad Science Journal مجلة بغداد للعلوم ISSN: 20788665 24117986 Year: 2010 Volume: 7 Issue: عدد خاص بمؤتمر العلمي النسوي 1 Pages: 411-414
Publisher: Baghdad University جامعة بغداد

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Abstract

Sex chromatin (SC) is defined as inactive X chromosome in the female somatic cells, it condensed as heterochromatic at interphase of the nucleus and appears like nuclear satellite projection, and only female cells contain SC in mammals. Different ages of cocks: immature (8,16 weeks old), growing (24 weeks old) and production age (32,48 weeks old) , were studied to detect the sex chromatin (SC) with it's various shapes :Sessile nodule(SS), Drum stick(DS), Tear drop(TD) and Club shape(CS) in the blood lymphocytes of blood smears prepared from WBCs of 45 New Hampshire(NH) strain cocks. Also SC measurements were detected which include: vertical (V) and horizontal (H) axes, SC area (SCA), nuclear area (NA), and the ratio(R) between these two areas. . Results show that SC is present in the male chicken blood lymphocytes in all ages. There is no significant difference in the incidence of the SC or its various measurements between ages, but the vertical axis was recorded a significant (P<0.05) decrease at 16 week old as compared with other ages. Also there is a non significant increase in the incidence of DS shape with ages; this indicated that simple, easy and cheap test could be helpful to detect some physiological parameters in the poultry species and other mammals.

يعرف الصبغين الجنسي بأنه احد كروموسومي X الذي فقد فعاليته في نواة الطور البيني للخلايا الجسمية في إناث اللبائن ويظهر كبروز متغاير الصبغين ملاصق لغشاء نواة الخلية . استخدمت في الدراسة أعمار مختلفة من الدجاج من سلالة النيوهمشاير : أعمار صغيرة (8و16 أسبوع), عمر النمو (24 أسبوع) وعمر البلوغ الجنسي (32و48 أسبوع ). تمت دراسة صفات الصبغين الجنسي وأشكاله المختلفة : عصا الطبال, بروز بدون ساق, دمعة العين والشكل الهراوي .تم العثور على هذه الأشكال في خلايا الدم البيضاء اللمفاوية من 45 ذكر من السلالة. تمت دراسة قياسات وأبعاد الصبغين الجنسي أيضا والتي تتضمن: أبعاد الصبغين الجنسي العمودي والأفقي ,مساحة الصبغين الجنسي ومساحة النواة الحاوية عليه والنسبة بينهما. أظهرت نتائج الدراسة وجود الصبغين الجنسي في انويه خلايا الدم اللمفاوية في ذكور الدجاج وفي جميع الأعمار بنسب متفاوتة. سجلت الدراسة عدم وجود أهمية معنوية في النسبة المئوية للصبغين الجنسي وأشكاله المختلفة أو أبعاده المختلفة بين مختلف الأعمار ,فقط البعد العمودي سجل انخفاضا معنويا (P<0.05) في عمر 16 أسبوعا مقارنة مع بقية الأعمار أيضا سجل ارتفاعا غير معنوي في نسبة تواجد شكل عصا الطبال مع تقدم العمر وهذا يشير انه من خلال فحص بسيط وسهل وغير مكلف يساعد على تحديد بعض الخواص الفسلجية المهمة في الدجاج واللبائن الأخرى.


Article
Determination of some Virulence Factors of Pasteurella multocia isolated from Human and Farm Animals infections
تحديد بعض عوامل الضراوة لبكتريا Pasteurella multocida المعزولة من إصابات الإنسان والحيوانات الحقلية

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Abstract

Twenty one Isolates of Pasteurella multocida were obtained from different clinical specimens of human and farm animals' infections. Human specimens included wounds swabs taken after cats and dog's bits beside the urine and sputum samples. The animal samples were nasal swabs and blood samples taken from chattels, also the poultry tissues of infected chicken with fowl cholera were collected. Bacterial isolates were isolated using Pasteurella multocida selective agar (PMSA) then identified doing different morphological, biochemical tests followed by api 20E diagnosis. The ability of the bacterial isolates to produce different virulence factors were studied, 18 isolates 85.7% were able to produce Lipase enzyme. Results of pathogenicity study on Lab. animals (mice) showed that there were 9 highly virulent isolates among the 21 (42.8%). The killing time was in between (10-24) hrs after injected the mice’s intraperitoneally. Some of the isolates showed their ability to produce the dermonecrotic toxin and the positive result appeared as highly purulent, dermonecrotic lesions after injection Genia pig intradermally. Results of antibiotic sensitivity test revealed that there were considerable variations in isolates susceptibility. Some of the isolates were highly resisted most of the used antibiotics while others were not. The crude bacterial extract from PMA20 isolate was tested to determine its ability to stimulate human lymphocytes division in vitro, results showed that the extract was able to stimulate cells division when the mitotic index was 1.2% as compared with the positive control.

أمكن الحصول على 21 عزلة من بكتريا Pasteurella multocida من نماذج مختلفة شملت مسحات من الجروح الناجمة عن عضات الكلاب والقطط وعينات القشع والادرار من الإنسان فيما شملت العينات الحيوانية أخذ مسحات الأنف وعينات من دم قطعان الماشية فضلا عن اخذ عينات من أنسجة دواجن ميتة مصابة بكوليرا الدجاج . عزلت البكتريا باستخدام وسط اكار الباستورلا-مالتوسيدا الانتخابي ثم شخصت اعتمادا على عدد من الفحوص المورفولوجية والبايوكيميائية فضلا عن التشخيص بنظام api 20E . تم التحري عن قابلية العزلات عن انتاج عدد من عوامل الضراوة اذ أظهرت النتائج قابلية 18 عزلة 85.7% على إنتاج الأنزيم المحلل للدهن Lipase كما بينت الدراسة أن هنالك 9 عزلات شديدة الضراوة للفئران من مجموع 21 (42.8%) ، وقد تراوح وقت الهلاك ما بين (24-10) ساعة بعد حقن الحيوانات بغشاء البريتون . استطاعت بعض من العزلات من إنتاج السموم المنخرة للجلد ، وظهرت النتيجة الموجبة من خلال حصول تقيح وتنخر شديدين بعد حقن خنازير غينيا تحت الجلد . فيما يخص نتائج فحص الحساسية للمضادات الحيوية فقد ظهر هنالك تفاوت في استجابة هذه العزلات اذ قاومت بعض منها وبشدة اغلب المضادات المستعملة في الوقت الذي استجابت بعض منها لهذه المضادات . اختبر المستخلص الخام للعزلة PMA20 للتحري عن قابليته لتحفيز خلايا دم الإنسان اللمفاوية على الانقسام في الزجاج اذ بينت النتائج قدرته المحفزه وبلغ معامل الانقسام %1.2 مقارنة مع نموذج السيطرة .


Article
Association between HLA-Class II Alleles and T-Cell Proliferation in Response to Enterovirus and Adenovirus

Author: Eman M. Saleh
Journal: Iraqi Academic Scientific Journal المجلة العراقية للاختصاصات الطبية ISSN: 16088360 Year: 2007 Volume: 6 Issue: 2 Pages: 102-108
Publisher: The Iraqi Borad for Medical Specialization المجلس العراقي للاختصاصات الطبية

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Abstract

BACK GROUND:Viruses may be involved in the pathogenesis of Type 1 Diabetes Mellitus (T1DM), either through direct β-cell infection or as triggers of autoimmunity.OBJECTIVE:To evaluate the T- cell proliferation in response to Enterovirus antigens including Coxsackievirus B and Poliovirus in addition to Adenovirus in an HLA- matched population of children with T1DM and children who were healthy.METHODS:A total of 60 Iraqi T1DM children were included in the presents study. They were new onset of the disease. For the purpose of comparisons, 50 apparently healthy control subjects were selected. HLA typing was measured by microlymphocytotoxicity, while methylthiazoltetrazolium (MTT) assay was used for lymphocyte proliferation by culturing peripheral blood lymphocytes (PBL) with Coxsackievirus B5, Adenovirus 3, 4, and 7, and Poliovaccin.RESULTS & CONCLUSION:No significant differences were shown in the PBL proliferative percentage in response to Con-A mitogen and tested viruses (CVB5 and Adenovirus) between T1DM and healthy controls, but PBL proliferative percentage of patients showed a significant decline in response to Poliovaccine. HLA class II (-DR3, DR4, DQ2 and DQ3) antigens were significantly increased in T1DM patients and they played an important role in the etiology of the disease. Strong T-cell proliferation in response to the tested viral antigens were observed to be related to HLA-DR4 and HLA-DQ3 antigens, whereas the HLA-DR3 and HLA-DQ2 alleles were associated with week responsiveness to the same antigens. However, in children with new- onset diabetes, responses were decreased and this could be caused by trapping of virus- specific T- cells in the pancreas


Article
New Methods for Establishment of Lymphoblastoid Cell Lines

Author: Ismail Ibrahim Latif
Journal: Diyala Journal For Pure Science مجلة ديالى للعلوم الصرفة ISSN: 83732222 25189255 Year: 2010 Volume: 6 Issue: 3 Pages: 277-287
Publisher: Diyala University جامعة ديالى

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Abstract

Background: Human lymphocytes can be isolated from whole blood by centrifugation using a commercially available high-density medium. This allows a single step gradient separation of blood, which yields the mononuclear cell ,the lymphocyte can be cultured specifically and will outgrow the others, eventually resulting in highly enriched population.Objective: this series of experiments were done as a trial to establish a new method for preparation of lymphocytes cell line.Methods: the same protocols were applied to all individuals, they were separated in to two groups:- Group 1:- one ml of heparinized blood were cultured in tissue culture flask containing 9 ml of stimulation medium and incubated in tissue culture flasks at 37ºC (4) . The flasks were subcultured at 37 OC in CO2 incubator every 3 days.Group 2:- Mononuclear cells were separated from whole blood, washed, counted, assessed for viability, then 1 ml of 2x106 cells were cultured in tissue culture flask containing stimulation medium and. The flask was incubated at 37 OC in CO2 incubator for 3 days. Another 1ml of mononuclear cells was cultured in tissue culture flask containing 1ml of growth medium, the flasks were incubated at 37 OC in CO2 incubator for 3 days.For both groups the cell cultures were propagated and maintained. Results: After initiation of cell culture for both groups, the mononuclear cell cultures were maintained up for 3 weeks, pure rich mononuclear cells were obtained and seen under inverted microscope.Conclusion: a new method for establishment of lymphoblast cell line were developed by cultivation of lymphocyte from whole blood . pure rich lymphocytes culture were obtained and maintained

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