이 작품은 코르크 사우스 의무실 빅토리아 대학 병원 유방 펀드, 아일랜드 암 협회 (CRI07TAN)와 코르크의 암 연구 센터를 통해 투자되었다. 안정 루시페라제을 표현 루이스 폐 세포 라인 박사에서 어떤 선물 Jooss 카린, 세포 Genesys, 주식 회사, 샌프란 시스코, 미국.

<html> 소개 간은 종종 전이성 질병의 첫 번째 사이트입니다 고급 질환 환자의 많은 40 30 %에서 확산의 유일한 사이트가 될 수 있습니다. 그러나 전이성 간 종양을 치료하기위한 옵션은 절제술에 적합한 환자의 소수로 몇 가지하고 chemotherapeutics에만 23% 주요 조치. 이 분야에 잠복기 연구는 절연 간 질환을 연구하기 위해 적합한 마우스 모델의 부재에 의해 제한되었습니다. 우리는 안정적으로 균일, 확산 간 metastases을 수립하는 데 사용할 수있는 분할 비장의 접근 방식을 설명합니다. 비장의 절반 절제술은 지라 종양의 성장과 비장의 다른 절반 지라 면역 보호 특성을 유지하기 위해 손상 포털 순환과 함께 반환되는의 결과를 피할 수. 우리가 설명하는 모델은 수행하기 위해 아주 간단하고 단일 절개를 통해 신속하게 수행할 수 있습니다. 종양 세포 라인의 발광 생물 특성은 동물의 희생없이 치료 반응의 정확한 양을 정함 수 있습니다. 프로토콜 I. 셀 준비 <ol><li> 루이스 폐 암종 세포는 37 10 % 태아 송아지 혈청 보충 Dulbecco의 수정된 필수 중간 ° C, 5 % CO 2를 증가하고 trypsinisation 때 하위 합류하여 수확하고 있습니다. </li><li> 세포 혈청 무료로 미디어에 준비되었습니다 Nucleocounter (화학 metec, 덴마크) 200 μl 1 X 10 6 세포의 단일 세포 현탁액을 사용하여 계산되었다. </li></ol> II. 마우스 준비 참고 : 모든 절차는 대학 코크의 윤리위원회에 의해 승인되었습니다. 저희 연구실에서는 마우스를 마취하는 주사용 anesthetics를 사용하여, 또는 하나는 동일한 효과를 달성하기위한 흡입 anesthetics을 사용할 수 있습니다. 우리가 사용하는 마우스 변형은 할런 연구소 (옥스 퍼 드셔, 영국)에서 얻은 사나이 답게 MF1 뉴 / 뉴 변형입니다. 16-22그램 연령 6-8 주간의 무게여 MF1 뉴 / 뉴 생쥐는이 실험에 포함되었습니다. 같은 C57와 Balb / C와 같은 다른 변종도 적절한 종양 세포 라인과 함께 사용할 수 있습니다. <ol><li> Anaesthesia는 100ul 1.5 MG의 케타민을 300 UG의 xylazine의 intraperitoneal 주사에 의해 전달되었다. </li><li> 마취의 깊이는 발가락 핀치를 사용하여 평가됩니다. 발가락 핀치와도 금단 반응이 없어야합니다. </li><li> anesthetized 마우스가 제대로 위치하고 손난로 지워졌을 것입니다. </li><li> 복부는 10 % betadine 솔루션 준비합니다. </li><li> 복부와 외과 사이트는 살균 방식에 draped 있습니다. </li></ol> III. 복부 절개 <ol><li> 작은 subcostal 절개는 피부에 이루어집니다. </li><li> 절개는 2~3cm로 연장됩니다. </li><li> 복벽 근육을 나누어 복막 캐비티가 입력되었습니다. </li></ol> IV. 노출과 비장 학부 <ol><li> 비장은 미리 절단 멸균 거즈에 동원하고 부드럽게 exteriorized입니다. </li><li> 두 4-0 vicryl ligatures는 비장의 중반 몸​​ 주위에 연결되어 있습니다. </li><li> 비장 두 헤미 - spleens을 만드는 두 ligatures 사이에 나눌 수 있습니다. </li></ol> 헤미 - 비장에 전지의 V. 사출 <ol><li> 27 G는 하나 헤미 - 비장에 세포 100 μL 볼륨을 삽입하는 데 사용됩니다. </li><li> Tumour 세포는 10 분 간 도달 주입된 세포의 대부분을위한 충분한 시간을 제공하는 간으로 배수하기 위해 사용할 수 있습니다. </li></ol> VI. 의 절단 헤미 - 비장, 복부 폐쇄 및 복구 <ol><li> 주입 헤미 - 비장이 excised됩니다. </li><li> 다른 헤미 - 비장은 복강에 반환됩니다. </li><li> 근육 계층은 간단한 실행 바늘을 사용하여 4-0 PDS의 봉합과 함께 닫힙니다. </li><li> 복부 피부가 중단 봉합사를 사용하여 4-0 Prolene와 함께 닫힙니다. </li><li> 유체 볼러스가 관리하고 이후 수술 진통제, Carpofen는 (5 밀리그램 / kg) 24 H 포스트 수술에 대한 IP 주입에 의해 관리됩니다. </li><li> 마우스가 마취에서 회복하기 위해 허용 </li></ol> VII. IVIS 전신 발광 생물 이미징 <ol><li> 마우스 주사용 마취제를 사용하여 anesthetized 있습니다. </li><li> Luciferin 기판은 이미징 전에 내부 복막 주사 십분에 의해 주어집니다 </li><li> 생쥐는 무대에 위치하고 IVIS 온 몸이 이미징 시스템을 사용하여 3 분 군데 아르 </li><li> 간장의 발광은 관심이 도구의 영역을 사용하여 측정할 수 </li></ol> VIII. 마우스 간의 노출 <ol><li> 마우스 주사용 마취제를 사용하여 anesthetized 있습니다. </li><li> 옥상 복부 절개는 생체내의 확산 간 전이의 시각화 수 있습니다. </li><li> 마우스는 자궁 전위에 의해 euthanized이며 간장은 추가 분석을 위해 excised 수 있습니다. </li></ol>_upload/1977/1977fig1.jpg &quot;고도는 =&quot;그림 1 &quot;/&gt; 그림 간은 1. 갓 excised murine 치료 및 종양 - 침입. <img src="/files/ftp_upload/1977/1977fig2.jpg" alt="그림 2" /> 그림 2.로서 IVIS 온 몸이 이미징에 의해 입증 간장 종양의 성장 속도론.

<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>

관심 없음 충돌 선언하지 않습니다.

간 전이의 연구에 대한 생체내 모델로 신뢰성의 개발은 새로운 치료 전략의 도래와 함께 중요한 증가되고있다. 새로운 전략의 정확한 연구를위한 동물 모델을 수행하기 위해 상대적으로 쉽게해야, 전이의 상당수를 형성 전이성 폭포의 모든 단계를 포함하고 전이성 세포의 제한된 수의의 감지 및 카운팅 수있는 기자 시스템을합니다. 로 설명한 분할 비장의 접근 방식은 단일 작업과 관련된...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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<tr>
<td>DMEM</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>D6429</td>
<td>&nbsp;</td>
<tr>
<td>PBS</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>D8537</td>
<td>&nbsp;</td>
<tr>
<td>Ketamine</td>
<td>&nbsp;</td>
<td>Vetoquinol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Xylazine</td>
<td>&nbsp;</td>
<td>Vetoquinol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Carprofen</td>
<td>&nbsp;</td>
<td>Vetoquinol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Videne</td>
<td>&nbsp;</td>
<td>Ecolab, Leeds, UK</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Vicryl 4-0 suture</td>
<td>&nbsp;</td>
<td>Ethicon</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>PDS 4-0 suture</td>
<td>&nbsp;</td>
<td>Ethicon</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Prolene 4-0 suture</td>
<td>&nbsp;</td>
<td>Ethicon</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Firefly Luciferin</td>
<td>&nbsp;</td>
<td>Biosynth, Zurich, Switzerland</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
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		</div>

murine bioluminescent hepatic tumour model

이 동영상은 이후 발광 생물 이미징으로 분석 수있는 마우스 모델에서 간 metastases의 설립을 설명합니다. Tumour 세포는 비장의 각 절반 작은 꽃자루 그대로 혈관을두고 두로 비장과 분할의 동원을 통해 지역화된 간 종양을 유발하는 간을 위해 특별히 관리하고 있습니다. constitutively 반딧불의 루시페라제 유전자 (luc1)를 표현 루이스 폐 암종 세포는 다음 10 분 후 resected 하나 헤미 - 비장에 주사를하고 있습니다. 다른 헤미 - 비장은 그대로 왼쪽 복부에 반환됩니다. 간 종양의 성장은 IVIS 온 몸이 이미징 시스템을 사용 bioluminescence 이미징에 의해 모니터할 수 있습니다. IVIS를 사용하여 종양 성장의 양적 이미징이 가능한 종양 세포의 정확한 quantitation를 제공합니다. Tumour 세포 죽음과 약물 치료로 인한 괴사는 발광 생물 신호의 감소에 의해 초기 표시됩니다. 이 마우스 모델은 전이성 종양 세포의 생존과 성장을 기본 메커니즘을 조사 가능하며 간 전이의 치료제의 평가에 사용될 수 있습니다.

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

Murine Bioluminescent Hepatic Tumour Model

이 문서는 생쥐에 orthotopic 발광 생물 간 종양의 유도, 그리고 라이브 온 몸이 발광 이미징을 사용하여 간장에 국한 종양 성장의 후속 분석을 위해 절차를 설명합니다.

Murine 발광 생물 간장 Tumour 모델

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

Research

JoVE Journal

Medicine

17.1K 조회수.  University College Cork. 이 문서는 생쥐에 orthotopic 발광 생물 간 종양의 유도, 그리고 라이브 온 몸이 발광 이미징을 사용하여 간장에 국한 종양 성장의 후속 분석을 위해 절차를 설명합니다.

비디오: 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.

인간 Mammary 섬유아 세포에서 암종 - 관련 섬유아 세포의 실험 세대 (CAFs)

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

연구

의학

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.

생체 듀얼 기판 발광 생물 이미징에

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.

이후 3D로 Intracranial 주입 VIVO에서</emMurine Gliomas의> 발광 생물 이미징

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.

Stereotactic Intracranial 주입 및<em&gt; 생체에</emGlioblastoma의 Multiforme의 마우스 모델 시스템에서 종양 Xenografts의&gt; 발광 생물 이미징

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.

상처 치유의 생쥐 모델

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.

췌장암의 진행 발광 동소 모델

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.

지주막 하 출혈의 생쥐 모델

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.

간 전이의 전임상 쥐과 모델

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.

쥐 간 문부 클램프 모델을 사용하여 직접 분절 인트라 간 납품 방법

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.

Myography 와이어를 사용 하 여 마우스 Mesenteric 동맥의 isometric 수축 측정

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

Methods Article

Murine 발광 생물 간장 Tumour 모델