وقد تم تمويل هذا العمل من خلال صندوق كورك جنوب مستشفى جامعة فيكتوريا في مستشفى الثدي ، وجمعية السرطان الأيرلندية (CRI07TAN) ومركز أبحاث السرطان كورك. وكان لويس خط الخلية الرئة معربا عن ستابلي luciferase هدية عينية من الدكتورة كارين Jooss ، خلية جينيسيس ، وشركة ، سان فرانسيسكو ، الولايات المتحدة الأمريكية.

<html> مقدمة الكبد وغالبا ما يكون أول موقع من المرض المنتشر ، وربما يكون الموقع الوحيد للانتشار في ما يصل الى 30 40 ٪ من المرضى الذين يعانون من المرض المتقدمة. لكن الخيارات المتاحة لعلاج أورام الكبد المنتشر قليلة مع أقلية من المرضى مناسبة لاستئصال وفقط 23 ٪ من المستجيبين الرئيسية مبحث المعالجة الكيميائية. وقد اقتصرت البحوث قبل السريرية في هذا المجال بسبب عدم وجود نموذج الفأر مناسبة لدراسة أمراض الكبد معزولة. وصفنا نهج تقسيم الطحال التي يمكن استخدامها لإنشاء موثوق موحدة ، الانبثاث الكبد انتشارا. استئصال نصف هذا الطحال يتجنب العواقب المترتبة على نمو الورم الطحال ، وعاد والنصف الآخر في الطحال مع تداول بوابة سليمة للحفاظ على خصائص وقائية مناعية الطحال. والنموذج الذي وصف بسيط للغاية لتنفيذ ويمكن القيام به بسرعة من خلال شق واحد. خصائص bioluminescent الورم خط الخلية يسمح الكمي الدقيق للالاستجابة للعلاج من دون تضحية من الحيوانات. بروتوكول أولا تحضير خلية <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> تزرع خلايا سرطان الرئة متوسطة لويس Dulbecco من الضروري تعديل تستكمل مع 10 ٪ مصل العجل الجنين عند 37 درجة مئوية ، 5 ٪ CO 2 وتحصد trypsinisation عندما شبه متموجة. </li><li style=";text-align:right;direction:rtl"> احصي الخلايا باستخدام Nucleocounter (الكيماوي metec ، الدنمارك) وتعليق خلية واحدة من الخلايا 1 X 6 10 في 200 ميكرولتر أعدت في وسائل الإعلام الحرة المصل. </li></ol> II. إعداد الماوس ملاحظة : تمت الموافقة على جميع الإجراءات من قبل لجنة الأخلاق من جامعة كوليدج كورك. في مختبرنا نستخدم حقن التخدير لتخدير الماوس ، بدلا من ذلك ، يمكن للمرء أن استخدام المسكنات استنشاقه لتحقيق التأثير نفسه. سلالة الماوس كنا هو أصلع MF1 نو / نو سلالة تم الحصول عليها من مختبرات هارلان (أوكسفوردشاير ، انكلترا). شملت الإناث MF1 نو / نو الفئران وزنها 16-22 غ و6-8 أسابيع من العمر في هذه التجربة. ويمكن أيضا سلالات أخرى مثل C57 وBALB / ج مناسبة يمكن استخدامها مع خطوط الخلايا السرطانية. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وتم تسليم التخدير عن طريق الحقن داخل الصفاق الكيتامين 100ul 1.5 ملغ و 300 ميكروغرام زيلازين. </li><li style=";text-align:right;direction:rtl"> ويتم تقييم عمق التخدير باستخدام قرصة أخمص قدميه. وينبغي أن يكون هناك منعكس الانسحاب مع قرصة أخمص قدميه. </li><li style=";text-align:right;direction:rtl"> يتوضع بشكل صحيح الماوس تخدير ، وسجلت أكثر من وسادة التدفئة. </li><li style=";text-align:right;direction:rtl"> وهيأهم البطن بمحلول betadine 10 ٪. </li><li style=";text-align:right;direction:rtl"> ورايات البطن وموقع الجراحة بطريقة عقيمة. </li></ol> III. شق البطن <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> يتم إجراء شق صغير في الجلد تحت الضلع. </li><li style=";text-align:right;direction:rtl"> ويمتد الشق إلى 2-3 سم. </li><li style=";text-align:right;direction:rtl"> تم تقسيم عضلات البطن الجدار ودخلت التجويف البريتوني. </li></ol> رابعا. وتعرض شعبة الطحال <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وحشدت الطحال وexteriorized بلطف على شاش معقم قبل قطع. </li><li style=";text-align:right;direction:rtl"> وترتبط اثنين من الحروف المركبة vicryl 4-0 في جميع أنحاء الجسم منتصف الطحال. </li><li style=";text-align:right;direction:rtl"> وينقسم الطحال بين الحروف المركبة two two خلق نصفي الطحال. </li></ol> خامسا حقن الخلايا في نصفي الطحال <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وتستخدم مجموعة من 27 إلى 100 وحدة ضخ ميكرولتر من الخلايا في واحد نصفي الطحال. </li><li style=";text-align:right;direction:rtl"> ويسمح للخلايا الأورام لنزيف في الكبد لمدة عشر دقائق وقتا كافيا لتوفير غالبية حقن الخلايا للوصول إلى الكبد. </li></ol> سادسا. استئصال الطحال ، هيمي ، وإغلاق البطن والاسترداد <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> يستأصل فيه حقن نصفي الطحال. </li><li style=";text-align:right;direction:rtl"> عاد هو الآخر نصفي الطحال إلى تجويف البطن. </li><li style=";text-align:right;direction:rtl"> إغلاق طبقة العضلات مع خياطة PDS 4-0 باستخدام غرزة بسيطة التشغيل. </li><li style=";text-align:right;direction:rtl"> إغلاق جلد البطن مع Prolene 4-0 باستخدام غرز توقف. </li><li style=";text-align:right;direction:rtl"> يدار بلعة السوائل وإدارة ما بعد الجراحة للوعي ، Carpofen (5 ملغ / كلغ) عن طريق الحقن في مجال الملكية الفكرية لمدة 24 ساعة بعد الجراحة. </li><li style=";text-align:right;direction:rtl"> ولا يسمح للماوس على التعافي من التخدير </li></ol> سابعا. IVIS كامل الجسم Bioluminescent التصوير <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> الفأر هو تخدير باستخدام حقن مخدر. </li><li style=";text-align:right;direction:rtl"> وتعطى Luciferin الركيزة التي داخل الصفاق عشر دقائق قبل الحقن التصوير </li><li style=";text-align:right;direction:rtl"> وضعت الفئران على المسرح وتصوير لمدة 3 دقائق باستخدام IVIS تصوير الجسم نظام الجامعة </li><li style=";text-align:right;direction:rtl"> ويمكن قياس التلألؤ من الكبد باستخدام أداة منطقة الفائدة </li></ol> ثامنا. التعرض للكبد الفأر <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> الفأر هو تخدير باستخدام حقن مخدر. </li><li style=";text-align:right;direction:rtl"> هناك شق في البطن على السطح يسمح التصور من ورم خبيث في الكبد المنتشر في الجسم الحي. </li><li style=";text-align:right;direction:rtl"> هو الموت الرحيم الماوس بواسطة الخلع عنق الرحم ، ويمكن استئصاله الكبد لإجراء مزيد من التحليل. </li></ol>_upload/1977/1977fig1.jpg &quot;بديل =&quot; الشكل 1 &quot;/&gt; الشكل 1. علاج الفئران يستأصل الورم طازجة وتسللت - الأكباد. <img src="/files/ftp_upload/1977/1977fig2.jpg" alt="الشكل 2" /> الشكل 2. حركية نمو الورم الكبدي كما يتضح من التصوير IVIS الجسم كله.

<ol>
	<li>Kasuya, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+subcutaneous+spleen+reservoir+for+multiple+portal+venous+injections+to+treat+liver+malignancies.">Mouse models of subcutaneous spleen reservoir for multiple portal venous injections to treat liver malignancies.</a> Cancer Res. 65, 3823-3827 (2005).</li><li>Liu, Q. Z., Tuo, C. W., Wang, B., Wu, B. Q., Zhang, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+metastasis+models+of+human+colorectal+carcinoma+established+in+nude+mice+by+orthotopic+transplantation+and+their+biologic+characteristic.">Liver metastasis models of human colorectal carcinoma established in nude mice by orthotopic transplantation and their biologic characteristic.</a> World J Gastroenterol. 4, 409-411 (1998).</li><li>Li, Z. Y., Ni, S., Yang, X., Kiviat, N., Lieber, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Xenograft+models+for+liver+metastasis:+Relationship+between+tumor+morphology+and+adenovirus+vector+transduction.">Xenograft models for liver metastasis: Relationship between tumor morphology and adenovirus vector transduction.</a> Mol Ther. 9, 650-657 (2004).</li><li>Cashman, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+gene+therapy+as+a+neoadjuvant+to+surgical+excision+to+control+metastatic+cancers.">Immune gene therapy as a neoadjuvant to surgical excision to control metastatic cancers.</a> Cancer Lett. , (2008).</li></ol>

الإعلان عن أي تضارب في المصالح.

وضع موثوق بها في النماذج الحية لدراسة الكبد الانبثاث أصبحت متزايدة الأهمية مع ظهور استراتيجيات علاجية جديدة. لدراسة دقيقة لوضع استراتيجيات جديدة ونماذج حيوانية ينبغي أن يكون من السهل نسبيا على أداء وشكل عددا كبيرا من ورم خبيث ، وإشراك جميع الخطوات من تتال?...

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		<th>Comment</th>
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<td>DMEM</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>D6429</td>
<td>&nbsp;</td>
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<td>PBS</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>D8537</td>
<td>&nbsp;</td>
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<td>Ketamine</td>
<td>&nbsp;</td>
<td>Vetoquinol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Xylazine</td>
<td>&nbsp;</td>
<td>Vetoquinol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Carprofen</td>
<td>&nbsp;</td>
<td>Vetoquinol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Videne</td>
<td>&nbsp;</td>
<td>Ecolab, Leeds, UK</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Vicryl 4-0 suture</td>
<td>&nbsp;</td>
<td>Ethicon</td>
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<td>PDS 4-0 suture</td>
<td>&nbsp;</td>
<td>Ethicon</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Prolene 4-0 suture</td>
<td>&nbsp;</td>
<td>Ethicon</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Firefly Luciferin</td>
<td>&nbsp;</td>
<td>Biosynth, Zurich, Switzerland</td>
<td>&nbsp;</td>
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murine bioluminescent hepatic tumour model

هذا الفيديو يصف إنشاء الانبثاث الكبد في نموذج الفأر التي يمكن تحليلها لاحقا من قبل bioluminescent التصوير. تدار خلايا الورم على وجه التحديد إلى الكبد للحث على وجود ورم في الكبد المترجمة ، عن طريق تعبئة والطحال وتقسيمه الى قسمين ، وترك الحبل على الغارب في الأوعية الدموية عنيق لكل نصف الطحال. يتم تلقيح خلايا سرطان الرئة لويس التي تعبر عن هذا الجين constitutively luciferase يراعة (luc1) إلى واحد نصفي الطحال وهي مقطوعة ثم 10 دقيقة في وقت لاحق. ويترك الآخر نصفي الطحال سليمة ، وعاد إلى البطن. ويمكن رصد نمو الورم في الكبد بواسطة التصوير تلألؤ بيولوجي باستخدام التصوير IVIS نظام الجسم كله. التصوير الكمي لنمو الورم باستخدام IVIS يوفر الكميات الدقيقة من الخلايا السرطانية قابلة للحياة. يشار إلى موت الخلايا والأورام نخر بسبب العلاج من تعاطي المخدرات في وقت مبكر عن طريق تخفيض في إشارة bioluminescent. هذا النموذج يسمح الماوس عن التحقيق في الآليات الكامنة الورم النقيلي خلايا البقاء والنمو ، ويمكن استخدامها لتقييم علاجات من ورم خبيث في الكبد.

Watch this Scientific Journal Video about Murine Bioluminescent Hepatic Tumour Model at JoVE.com

Murine Bioluminescent Hepatic Tumour Model

توضح هذه المقالة إجراء لتحريض مثلي bioluminescent أورام الكبد في الفئران ، وتحليل لاحقة من نمو الورم محصورا في الكبد باستخدام يعيش التلألؤ الجسم كله التصوير.

فأري الورم الكبدي Bioluminescent النموذجي

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are ...

murine-bioluminescent-hepatic-tumour-model

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17.1K Views.  University College Cork. توضح هذه المقالة إجراء لتحريض مثلي bioluminescent أورام الكبد في الفئران ، وتحليل لاحقة من نمو الورم محصورا في الكبد باستخدام يعيش التلألؤ الجسم كله التصوير.

Video: Murine Bioluminescent Hepatic Tumour Model

Experimental Generation of Carcinoma-Associated Fibroblasts (CAFs) from Human Mammary Fibroblasts

Carcinomas are complex tissues comprised of neoplastic cells and a non-cancerous compartment referred to as the 'stroma'. The stroma consists of extracellular matrix (ECM) and a variety of mesenchymal cells, including fibroblasts, myofibroblasts, endothelial cells, pericytes and leukocytes 1-3.
The tumour-associated stroma is responsive to substantial paracrine signals released by neighbouring carcinoma cells. During the disease process, the stroma often becomes populated by carcinoma-associated fibroblasts (CAFs) including large numbers of myofibroblasts. These cells have previously been extracted from many different types of human carcinomas for their in vitro culture. A subpopulation of CAFs is distinguishable through their up-regulation of &alpha;-smooth muscle actin (&alpha;-SMA) expression4,5. These cells are a hallmark of 'activated fibroblasts' that share similar properties with myofibroblasts commonly observed in injured and fibrotic tissues 6. The presence of this myofibroblastic CAF subset is highly related to high-grade malignancies and associated with poor prognoses in patients.
Many laboratories, including our own, have shown that CAFs, when injected with carcinoma cells into immunodeficient mice, are capable of substantially promoting tumourigenesis 7-10. CAFs prepared from carcinoma patients, however, frequently undergo senescence during propagation in culture limiting the extensiveness of their use throughout ongoing experimentation. To overcome this difficulty, we developed a novel technique to experimentally generate immortalised human mammary CAF cell lines (exp-CAFs) from human mammary fibroblasts, using a coimplantation breast tumour xenograft model.
In order to generate exp-CAFs, parental human mammary fibroblasts, obtained from the reduction mammoplasty tissue, were first immortalised with hTERT, the catalytic subunit of the telomerase holoenzyme, and engineered to express GFP and a puromycin resistance gene. These cells were coimplanted with MCF-7 human breast carcinoma cells expressing an activated ras oncogene (MCF-7-ras cells) into a mouse xenograft. After a period of incubation in vivo, the initially injected human mammary fibroblasts were extracted from the tumour xenografts on the basis of their puromycin resistance 11.
We observed that the resident human mammary fibroblasts have differentiated, adopting a myofibroblastic phenotype and acquired tumour-promoting properties during the course of tumour progression. Importantly, these cells, defined as exp-CAFs, closely mimic the tumour-promoting myofibroblastic phenotype of CAFs isolated from breast carcinomas dissected from patients. Our tumour xenograft-derived exp-CAFs therefore provide an effective model to study the biology of CAFs in human breast carcinomas. The described protocol may also be extended for generating and characterising various CAF populations derived from other types of human carcinomas.

Experimental Generation of Carcinoma-Associated Fibroblasts CAFs from Human Mammary Fibroblasts

Carcinoma-associated fibroblasts (CAFs) rich in myofibroblasts present within the tumour stroma, play a major role in driving tumour progression. We developed a coimplantation tumour xengraft model for experimentally generating CAFs from human mammary fibroblasts. The protocol describes how to establish CAF myofibroblasts that acquire an ability to promote tumourigenesis.

الجيل التجريبية للسرطان ، المرتبطين بها الخلايا الليفية (مقاهي) من الخلايا الليفية الثديية البشرية

Carcinomas are complex tissues comprised of neoplastic cells and a non-cancerous compartment referred to as the 'stroma'. The stroma consists of ...

In vivo Dual Substrate Bioluminescent Imaging

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate since the advent of in vivo bioluminescent imaging technologies 1-3. Indeed, the ability to locate and quantify tumor growth longitudinally in a single cohort of animals to completion of the study as opposed to sacrificing individual groups of animals at specific assay times has revolutionized how researchers investigate breast cancer metastasis. Unfortunately, current methodologies preclude the real-time assessment of critical changes that transpire in cell signaling systems as breast cancer cells (i) evolve within primary tumors, (ii) disseminate throughout the body, and (iii) reinitiate proliferative programs at sites of a metastatic lesion. However, recent advancements in bioluminescent imaging now make it possible to simultaneously quantify specific spatiotemporal changes in gene expression as a function of tumor development and metastatic progression via the use of dual substrate luminescence reactions. To do so, researchers take advantage for two light-producing luciferase enzymes isolated from the firefly (Photinus pyralis) and sea pansy (Renilla reniformis), both of which react to mutually exclusive substrates that previously facilitated their wide-spread use in in vitro cell-based reporter gene assays 4. Here we demonstrate the in vivo utility of these two enzymes such that one luminescence reaction specifically marks the size and location of a developing tumor, while the second luminescent reaction serves as a means to visualize the activation status of specific signaling systems during distinct stages of tumor and metastasis development. Thus, the objectives of this study are two-fold. First, we will describe the steps necessary to construct dual bioluminescent reporter cell lines, as well as those needed to facilitate their use in visualizing the spatiotemporal regulation of gene expression during specific steps of the metastatic cascade. Using the 4T1 model of breast cancer metastasis, we show that the in vivo activity of a synthetic Smad Binding Element (SBE) promoter was decreased dramatically in pulmonary metastasis as compared to that measured in the primary tumor 4-6. Recently, breast cancer metastasis was shown to be regulated by changes within the primary tumor microenvironment and reactive stroma, including those occurring in fibroblasts and infiltrating immune cells 7-9. Thus, our second objective will be to demonstrate the utility of dual bioluminescent techniques in monitoring the growth and localization of two unique cell populations harbored within a single animal during breast cancer growth and metastasis.

In vivo Dual Substrate Bioluminescent Imaging

Herein we describe the methods to construct, visualize, and quantify the bioluminescent reactions of both firefly and renilla luciferase enzymes expressed in metastatic breast cancer cells during their growth and metastasis in vivo.

في الجسم الحي التصوير المزدوج الركيزة Bioluminescent

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate ...

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The cell line was originally induced by intracranial injection of 3-methyl-cholantrene into a C57BL/6 syngeneic mouse strain 1; therefore, immunologically competent C57BL/6 mice can be used. While we use GL261, the following protocol can be used for the implantation and monitoring of any intracranial mouse tumor model. GL261 cells were engineered to stably express firefly luciferase (GL261-luc). We also created the brighter GL261-luc2 cell line by stable transfection of the luc2 gene expressed from the CMV promoter. C57BL/6-cBrd/cBrd/Cr mice (albino variant of C57BL/6) from the National Cancer Institute, Frederick, MD were used to eliminate the light attenuation caused by black skin and fur. With the use of albino C57BL/6 mice; in vivo imaging using the IVIS Spectrum in vivo imaging system is possible from the day of implantation (Caliper Life Sciences, Hopkinton, MA). The GL261-luc and GL261-luc2 cell lines showed the same in vivo behavior as the parental GL261 cells. Some of the shared histological features present in human GBMs and this mouse model include: tumor necrosis, pseudopalisades, neovascularization, invasion, hypercellularity, and inflammation 1.
Prior to implantation animals were anesthetized by an intraperitoneal injection of ketamine (50 mg/kg), xylazine (5 mg/kg) and buprenorphine (0.05 mg/kg), placed in a stereotactic apparatus and an incision was made with a scalpel over the cranial midline. A burrhole was made 0.1mm posterior to the bregma and 2.3mm to the right of the midline. A needle was inserted to a depth of 3mm and withdrawn 0.4mm to a depth of 2.6mm. Two &mu;l of GL261-luc or GL261-luc2 cells (107 cells/ml) were infused over the course of 3 minutes. The burrhole was closed with bonewax and the incision was sutured.
Following stereotactic implantation the bioluminescent cells are detectable from the day of implantation and the tumor can be analyzed using the 3D image reconstruction feature of the IVIS Spectrum instrument. Animals receive a subcutaneous injection of 150&mu;g luciferin /kg body weight 20 min prior to imaging. Tumor burden is quantified using mean tumor bioluminescence over time. Tumor-bearing mice were observed daily to assess morbidity and were euthanized when one or more of the following symptoms are present: lethargy, failure to ambulate, hunched posture, failure to groom, anorexia resulting in &gt;10% loss of weight. Tumors were evident in all of the animals on necropsy.

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

Intracranial implantation of GL261 cells into C57BL/6 mice produces malignant gliomas that recapitulate many of the hallmarks of human glioblastoma multiforme. We used GL261 cells stably expressing luciferase to allow us to use in vivo imaging to follow tumor progression. The surgery and 3D in vivo imaging are demonstrated.

زرع داخل القحف مع 3d اللاحقة في فيفو Bioluminescent التصوير الاورام الدبقية الفئران

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The ...

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality treatment consisting of surgical resection, post-operative radiation therapy and temozolomide chemotherapy 1. New therapeutic approaches are clearly needed to improve patient survival and quality of life. The development of more effective treatment strategies would be aided by animal models of GBM that recapitulate human disease yet allow serial imaging to monitor tumor growth and treatment response. In this paper, we describe our technique for the precise stereotactic implantation of bio-imageable GBM cancer cells into the brains of nude mice resulting in tumor xenografts that recapitulate key clinical features of GBM 2. This method yields tumors that are reproducible and are located in precise anatomic locations while allowing in vivo bioluminescent imaging to serially monitor intracranial xenograft growth and response to treatments 3-5. This method is also well-tolerated by the animals with low perioperative morbidity and mortality.

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

We describe an integrated method for the precise, stereotactic implantation of human glioblastoma multiforme cells into the brains of nude mice and subsequent serial in vivo imaging to monitor growth and response to treatment of the resultant xenografts.

داخل الجمجمة وزراعة المجسم<em&gt; في الجسم الحي</em&gt; التصوير Bioluminescent من الأعضاء المستمدة من الحيوانات ورم في النظام النموذجي ماوس من الأشكال ورم أرومي دبقي

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

نموذج الفئران من التئام الجروح

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

نموذج مثلي إضاءة الحيوية من سرطان البنكرياس التقدم

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

A نموذج من الفئران تحت العنكبوتية نزف

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

وما قبل السريرية الفئران نموذج من الانبثاث الكبدي

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

طريقة التسليم المباشر القطاعي البينية الكبد باستخدام نموذج الفأر الكبد نقيري المشبك

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

متساوي القياس كونتراكتيليتي القياس الشريان المساريقي العلوي الماوس باستخدام أسلاك ميوجرافي

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...