Name: intracranial implantation with subsequent 3d in vivo bioluminescent imaging of murine gliomas
Uploaded: 2011-11-06T14:00:00.000+00:00
Duration: 9 min 46 s
Description: Watch this Scientific Journal Video about Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas at JoVE.com

우리는 GL261 룩 세포의 준비에 도움이 제안에 대한 lentivirus 시스템뿐만 아니라 Mahil 라오에 대한 plasmids의 관대한 선물에 대한 박사 조슈아 B. 루빈을 인정하고 싶습니다. 우리는 뇌 종양 연구 (SSBTR), 배로우 신경 재단과 관대한 지원 월러스 재단을 지원하는 학생 주셔서 감사합니다. 동물 실험은 세인트 조셉 병원과 의료 센터의 기관 동물 케어 및 사용위원회에 의해 명시된 지침과 규정에 따라 수행되었다.

<ol>
	<li>Candolfi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracranial+glioblastoma+models+in+preclinical+neuro-oncology:+neuropathological+characterization+and+tumor+progression.">Intracranial glioblastoma models in preclinical neuro-oncology: neuropathological characterization and tumor progression.</a> J Neurooncol. 85, 133-148 (2007).</li><li>Stafford, P., Abdelwahab, M. G., do, K. i. m., Preul, Y., Rho, M. C., M, J., Scheck, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+ketogenic+diet+reverses+gene+expression+patterns+and+redudes+reactive+oxygen+species+levels+when+used+as+an+adjuvant+therapy+for+glioma.">The ketogenic diet reverses gene expression patterns and redudes reactive oxygen species levels when used as an adjuvant therapy for glioma.</a> Nutr Metab. 7, (2010).</li><li>Caliper Life Sciences, Inc. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Living Image Software Version 4.0. VivoVision Systems. , (2010).</li><li>Paxinos, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Mouse Brain in Stereotaxic Coordinates. , (2001).</li><li>Jouanneau, E., Poujol, D., Gulia, S., Le, M. I., Blay, J. Y., Belin, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dendritic+cells+are+essential+for+priming+but+inefficient+for+boosting+antitumour+immune+response+in+an+orthotopic+murine+glioma+model.">Dendritic cells are essential for priming but inefficient for boosting antitumour immune response in an orthotopic murine glioma model.</a> Cancer Immunol Immunother. 55, 254-267 (2006).</li></ol>

이 기사에 무료로 액세스 캘리퍼스 생명 과학 후원됩니다.

셀 inoculum은 0.4 mm의 포켓을 만든 후 두뇌의 표면에서 2.6 mm의 깊이에 들어갈 수 있습니다. 바늘의 적절한 배치와 심도를 보장하기 위해 X - 레이는 C - 암 또는 이와 유사한 X - 선 이미지 가속화 장치를 사용 취할 수 있지만, 이것은 선택 사항입니다. 동물이 제대로 진정되지 않으면 수술의 합병증은 동물 세포 주입하는 동안 이동할 수있는 시점에서 발생할 수 있습니다. 이것은 바늘 트랙에서 셀 믹스 또는 출혈의 누출을 일으킬 수 있습니다. 세포의 누수는 종양 세포의 성장을 이소성됩니다. burrhole이 프로토콜 4 설명한 2.3 mm까지 중간 만든 경우에도 소중히 할 수있는 뇌실를 중요하지 않습니다. 바늘과 세포 확산의 적절한 배치는 2 μl 메틸렌 블루 염색와 마우스를 일으키는 및 주입 염료의 위치를​​ 확인하기 위해 뇌 조직을 해부하여 테스트되었습니다. 이 프로토콜에서 우리는 생체내 이미징 시스템 및 t의 IVIS 스펙트럼을 사용한그는이 악기와 함께 사용하도록 설계 이미지 소프트웨어 (V 4.0) 생활. (캘리퍼스 생명 과학). 생체내 이미징 시스템과 이미지 분석 도구의 모든 비교는 유사한 결과를 얻을하는 데 사용할 수 있습니다. 이러한 시스템은 실험 intracranial 종양의 성장에 따라 기존의 자기 공명 이미징 (MRI)을 통해 몇 가지 이점을 제공합니다. 가장 확실한는 2 악기의 상대적인 비용이다 - 동물 MRI 기계는 훨씬 더 비싼 있으며, 일반적으로 숙련된 기술자 MRI의 서비스를 필요로합니다. 이러한 여기에서 설명한 있었는지와 같은 생체내 이미징의 최종 사용자에 의해 할 수 있습니다. MRI의 데이터 quantitation는 많은 시간과 다소 엄밀하지 않은 상태에서 bioluminescence 데이터는 양적 있습니다. 또한, MRI 이미지는 부종과 종양 세포 이외에 염증을 표시하고 치료 효과에서 종양을 구분하기 어려울 수 있습니다. 이러한 이유로 성장하는 종양의 정확한 용적 측정을 획득하는 것은 도전이 될 수 있습니다. Bioluminescence는 ATP가 필요합니다그러므로 오직 살아있는 종양 세포는 종양의 크기 데이터에 기여하고 있습니다. 기계가 사용할 수있는 경우에도 불구하고, MRI를 몇 가지 이점이 있습니다. 세포는 MRI에 의해 시각이 될 수있는 발광 생물 마커로 표시 될 필요가 없습니다. 사망 시에 tumoral 부종을 시각화하는 능력은 몇 가지 실험 프로토콜에 대한 이점 수 있습니다. 두 기술을 사용할 때 두 기술의 강도가 하나를 사용하면 다른의 사용을 배제하지 않으므로 데이터는 종양의 성장뿐만 아니라 같은 동물의 사망 시에 tumoral 부종과 염증의 존재를 얻을 수 있습니다 연구원 수 있습니다.

<table border="1"><tbody><tr><td> 시약이나 공급의 명칭 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> GL261 - luc2 Bioware 울트라 </td><td> 캘리퍼스 생명 과학 </td><td> GL261 - luc2 </td><td></td></tr><tr><td> Dulbecco의 수정 독수리 중간 (DMEM) </td><td> Invitrogen </td><td> 10313039 </td><td></td></tr><tr><td> Geneticin (G418) </td><td> Gibco (Invitrogen) </td><td> 11811-023 </td><td></td></tr><tr><td> 소태아 세럼 (FCS) </td><td> Invitrogen </td><td> 26140079 </td><td></td></tr><tr><td> Phospate는 버​​퍼 식염수 (PBS) </td><td> Invitrogen </td><td> 70011044 </td><td></td></tr><tr><td> C57BL/6-cBrd/cBrd/Cr 마우스 </td><td> NCI - 프레드릭 </td><td></td><td></td></tr><tr><td> AKWA의 눈물 윤활유 안과 연고 </td><td> Akorn Inc의 </td><td> 17478-062-35 </td><td></td></tr><tr><td> Ketaset (케타민을 하이드로 클로라이드) </td><td> Wyeth </td><td> 11570775 </td><td></td></tr><tr><td> Sedazine (xylazine 염산염) </td><td> Wyeth </td><td> 10031894 </td><td></td></tr><tr><td> 작은 동물 Stereotaxic 악기 </td><td> Kopf 인 스트 루먼트 </td><td> 900 </td><td></td></tr><tr><td> SYS - Micro4 컨트롤러 UltraMicroPump </td><td> 세계 정밀 계측기 </td><td> UMP3 - 1 </td><td> 사용할 수없는 경우 수동으로 달이다 수 있습니다 </td></tr><tr><td> 26 게이지와 10μl 주사기는 바늘 beveled </td><td> 세계 정밀 계측기 </td><td> SGE010RNS </td><td></td></tr><tr><td> Adison 집게 </td><td> 세계 정밀 계측기 </td><td> 500,092 </td><td></td></tr><tr><td> Penfield 해부학자 </td><td> Codman </td><td> 65-1015 </td><td></td></tr><tr><td> 16g 1 ½ 정밀 글라이드 니들 </td><td> Beckton, 디킨슨과 회사 (BD) </td><td> 305,198 </td><td></td></tr><tr><td> 외과 블레이드 핸들 </td><td> BD </td><td> 371,030 </td><td></td></tr><tr><td> 15 사이즈 블레이드 </td><td> BD </td><td> 371,315 </td><td></td></tr><tr><td> 4-0 Vicryl 봉합사 </td><td> Ethicon </td><td> VCP496G </td><td></td></tr><tr><td> 뼈 왁스 </td><td> MEDLINE </td><td> DYNJBW25 </td><td></td></tr><tr><td> Povidine - 요오드 면봉 스틱 </td><td> MEDLINE </td><td> MD93901 </td><td></td></tr><tr><td> D - Luciferin의 칼륨 소금 </td><td> 캘리퍼스 생명 과학 </td><td> 122,796 </td><td></td></tr><tr><td> Forane (Isoflurane) </td><td> 벡스터 </td><td> 1001936060 </td><td></td></tr><tr><td> OPMI Pentero 현미경 </td><td> 칼 자이스 혈구, 주식 회사 </td><td></td><td> 모든 수술 현미경은 충분 </td></tr><tr><td> 선택 마취 시스템과 Xenogen IVIS 스펙트럼 </td><td> 캘리퍼스 생명 과학 </td><td></td><td></td></tr></tbody></table>

intracranial implantation with subsequent 3d in vivo bioluminescent imaging of murine gliomas

마우스 glioma 261 (GL261)은 생체내 모델 시스템에으로 인식되고 인간 glioblastoma multiforme의 (GBM)의 기능 중 많은 recapitulates. 셀 라인은 원래 C57BL / 6 마우스 syngeneic 스트레인 하나에 3 - 메틸 - cholantrene의 intracranial 주입에 의해 유도되었다, 따라서 immunologically 유능한 C57BL / 6 생쥐를 사용할 수 있습니다. 우리가 GL261을 사용하는 동안 다음과 같은 프로토콜은 어떤 intracranial 마우스 종양 모델의 주입 모니터링에 사용할 수 있습니다. GL261 세포는 안정적으로 표현 반딧불 루시페라제 (GL261 룩)으로 설계되었습니다. 우리는 또한 CMV 프로 모터에서 표현 luc2 유전자의 안정적인 transfection하여 밝은 GL261 - luc2 세포주를 만들었습니다. 국립 암 연구소, 프레데릭, MD에서 C57BL/6-cBrd/cBrd/Cr 마우스 (C57BL / 6의 흰둥이 변종)는 검은 피부와 모피로 인해 빛을 감쇠를 제거하는 데 사용되었습니다. 흰둥이 C57BL / 6 마우스의 사용을 통해, 생체내 이미징의 생체내 메신저에서 IVIS 스펙트럼을 사용하여시스템을 노화 것은 주입의 날 (캘리퍼스 생명 과학, Hopkinton, MA)에서 가능합니다. GL261 룩과 GL261 - luc2 셀 라인은 부모 GL261 세포로 생체내 행동 동일하게 나타났다. 인간 GBMs에있는 공유 histological 기능이 마우스 모델 중 일부는 다음과 같습니다 : 종양 괴사, pseudopalisades, neovascularization, 침공, hypercellularity과 염증 1. 이전 주입 동물 stereotactic기구와 절개에 배치 300ml의 intraperitoneal 주사 (50 MG / kg), xylazine (5 밀리그램 / kg)과 buprenorphine (0.05 밀리그램 / kg)에 의해 anesthetized되었다이 이상 메스로 만들어진 두개골 중간선. burrhole는 bregma과 중간선의 오른쪽에있는 2.3mm로 0.1mm 사후되었다. 바늘은 2.6mm의 깊이 3mm와 0.4mm 철회의 깊이로 삽입되었다. GL261 룩이나 GL261 - luc2 전지 (10 7 셀 / ML)의 두 μl는 삼분의 과정을 통해 주입했다. burrhole는 폐쇄되었습니다bonewax과 절개는 봉합했습니다와 함께. stereotactic 주입 따라 발광 생물 세포가 이식 당일부터 감지하고 종양이 IVIS 스펙트럼 악기의 3D 이미지 재구성 기능을 사용하여 분석하실 수 있습니다. 동물 150μg luciferin / 이전 이미지로 kg의 체중 20 분의 피하 주사를 받게됩니다. 종양 부담 시간이 지남에 따라 의미 종양의 bioluminescence를 사용하여 계량입니다. 종양 베어링 생쥐는 병적 상태를 평가하기 위해 매일 관찰되었으며 다음과 같은 현상이 하나 이상 존재하는 경우 euthanized되었습니다 혼수, ambulate에 실패, hunched 자세, 신랑에 실패, 거식증은 체중의&gt; 10 %의 손실 발생. 종양 검시에있는 동물의 모든 분명했다.

Watch this Scientific Journal Video about Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas at JoVE.com

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

C57BL에 GL261 셀 / 6 마​​우스의 Intracranial 이식은 인간 glioblastoma multiforme의의 특징 중 많은 것들을 요점을 되풀이하다 악성 gliomas을 생산하고 있습니다. 우리는 우리가 사용할 수 있도록 안정 루시페라제 표현 GL261 세포를 사용<em&gt; 생체내에</em&gt; 영상은 종양 진행을 따르십시오. 수술 및 3D<em&gt; 생체내에</em&gt; 영상을 보여주 있습니다.

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 ...

intracranial-implantation-with-subsequent-3d-vivo-bioluminescent

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

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

20.4K 조회수.  Barrow Neurological Institute of St. Joseph’s Hospital and Medical Center. 中文

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

Establishing Intracranial Brain Tumor Xenografts With Subsequent Analysis of Tumor Growth and Response to Therapy using Bioluminescence Imaging

Transplantation models using human brain tumor cells have served an essential function in neuro-oncology research for many years. In the past, the most commonly used procedure for human tumor xenograft establishment consisted of the collection of cells from culture flasks, followed by the subcutaneous injection of the collected cells in immunocompromised mice. Whereas this approach still sees frequent use in many laboratories, there has been a significant shift in emphasis over the past decade towards orthotopic xenograft establishment, which, in the instance of brain tumors, requires tumor cell injection into appropriate neuroanatomical structures. Because intracranial xenograft establishment eliminates the ability to monitor tumor growth through direct measurement, such as by use of calipers, the shift in emphasis towards orthotopic brain tumor xenograft models has necessitated the utilization of non-invasive imaging for assessing tumor burden in host animals. Of the currently available imaging methods, bioluminescence monitoring is generally considered to offer the best combination of sensitivity, expediency, and cost. Here, we will demonstrate procedures for orthotopic brain tumor establishment, and for monitoring tumor growth and response to treatment when testing experimental therapies.

Luciferase-modified human brain tumor xenografts can be established intracranially in athymic mice, with subsequent monitoring of tumor growth and response to therapy using bioluminescence imaging. In combination with survival analysis, bioluminescence monitoring is an essential research tool for pre-clinical testing of therapies being considered for treating brain tumors.

Bioluminescence 이미징을 사용하여 종양 성장과 치료에 반응 이후 분석 Intracranial 뇌종양의 Xenografts 구축

Transplantation models using human brain tumor cells have served an essential function in neuro-oncology research for many years. In the past, the most ...

연구

신경과학

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1&alpha; (HIF-1&alpha;) to drive transcription of various genes. Therefore, we have used a HIF-1&alpha; reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1&alpha; bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1&alpha; binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1&alpha; bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

Bioluminescence imaging of hypoxia inducible factor-1&alpha; activity is applied to monitor intracranial tumor hypoxia development in a breast cancer brain metastasis mouse model.

유방암 뇌 전이의 종양 Hypoxia 역학 생체내의 Bioluminescence 이미징에서는 마우스 모델에서

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There ...

의학

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting immunodeficient mice subcutaneously with cancer cells and measuring the tumor mass after it becomes visible and palpable. Orthotopic injections of cancer cells aim to introduce the xenograft in the microenvironment that most closely resembles the tissue of origin of the tumor being studied. Brain cancer research requires intracranial injection of cancer cells to allow the tumor formation and analysis in the unique microenvironment of the brain. The in vivo imaging of intracranial xenografts monitors instantaneously the tumor mass of orthotopically engrafted mice. Here we report the use of fluorescence molecular tomography (FMT) of brain tumor xenografts. The cancer cells are first transduced with near infrared fluorescent proteins and then injected in the brain of immunocompromised mice. The animals are then scanned to obtain quantitative information about the tumor mass over an extended period of time. Cell pre-labeling allows for cost effective, reproducible, and reliable quantification of the tumor burden within each mouse. We eliminated the need for injecting imaging substrates, and thus reduced the stress on the animals. A limitation of this approach is represented by the inability to detect very small masses; however, it has better resolution for larger masses than other techniques. It can be applied to evaluate the efficacy of a drug treatment or genetic alterations of glioma cell lines and patient-derived samples.

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Orthotopic intracranial injection of tumor cells has been used in cancer research to study brain tumor biology, progression, evolution, and therapeutic response. Here we present fluorescence molecular tomography of tumor xenografts, which provides real-time intravital imaging and quantification of a tumor mass in preclinical glioblastoma models.

Vivo에서 이미징 세포종 Xenografts의 형광 분자 단층 촬영

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting ...

암 연구학

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 ...

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a novel platform that allows for direct long-term visualization of tissue structure changes intracranially. Imaging at a single cell resolution in a real-time fashion provides supplementary dynamic information beyond that provided by standard end-point histological analysis, which looks solely at 'snap-shot' cross sections of tissue.
Establishing this intravital imaging technique in fluorescent chimeric mice, we are able to image four fluorescent channels simultaneously. By incorporating fluorescently labeled cells, such as GFP+ bone marrow, it is possible to track the fate of these cells studying their long-term migration, integration and differentiation within tissue. Further integration of a secondary reporter cell, such as an mCherry glioma tumor line, allows for characterization of cell:cell interactions. Structural changes in the tissue microenvironment can be highlighted through the addition of intra-vital dyes and antibodies, for example CD31 tagged antibodies and Dextran molecules.
Moreover, we describe the combination of our ICW imaging model with a small animal micro-irradiator that provides stereotactic irradiation, creating a platform through which the dynamic tissue changes that occur following the administration of ionizing irradiation can be assessed.
Current limitations of our model include penetrance of the microscope, which is limited to a depth of up to 900 &mu;m from the sub cortical surface, limiting imaging to the dorsal axis of the brain. The presence of the skull bone makes the ICW a more challenging technical procedure, compared to the more established and utilized chamber models currently used to study mammary tissue and fat pads 5-7. In addition, the ICW provides many challenges when optimizing the imaging.

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

소설 고해상도<em&gt; 생체 내</em종양의 성장과 치료학에 두개 내 구조물의 동적 응답을 공부하기&gt; 이미징 기술

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

An Orthotopic Glioblastoma Mouse Model Maintaining Brain Parenchymal Physical Constraints and Suitable for Intravital Two-photon Microscopy

Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date.
Murine models of this pathology rely on the injection of a suspension of glioma cells into the brain parenchyma following incision of the dura-mater. Whereas the cells have to be injected superficially to be accessible to intravital two-photon microscopy, superficial injections fail to recapitulate the physiopathological conditions. Indeed, escaping through the injection tract most tumor cells reach the extra-dural space where they expand abnormally fast in absence of mechanical constraints from the parenchyma.
Our improvements consist not only in focally implanting a glioma spheroid rather than injecting a suspension of glioma cells in the superficial layers of the cerebral cortex but also in clogging the injection site by a cross-linked dextran gel hemi-bead that is glued to the surrounding parenchyma and sealed to dura-mater with cyanoacrylate. Altogether these measures enforce the physiological expansion and infiltration of the tumor cells inside the brain parenchyma. Craniotomy was finally closed with a glass window cemented to the skull to allow chronic imaging over weeks in absence of scar tissue development.
Taking advantage of fluorescent transgenic animals grafted with fluorescent tumor cells we have shown that the dynamics of interactions occurring between glioma cells, neurons (e.g. Thy1-CFP mice) and vasculature (highlighted by an intravenous injection of a fluorescent dye) can be visualized by intravital two-photon microscopy during the progression of the disease.
The possibility to image a tumor at microscopic resolution in a minimally compromised cerebral environment represents an improvement of current GBM animal models which should benefit the field of neuro-oncology and drug testing.

We have established a cortical orthotopic glioblastoma model in mice for intravital two-photon microscopy that recapitulates the biophysical constraints normally at play during the growth of the tumor. A chronic glass window replacing the skull above the tumor enables the follow-up of the tumor progression over time by two-photon microscopy.

소성을 Glioblastoma의 마우스 모델은 뇌 실질 물리적 제약 조건을 유지하고의 intravital 두 광자 현미경에 적합

Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date.
Murine models of this pathology ...

마우스에서 2P-IVM을 사용한 뇌종양의 나노 입자 실시간 추적

Two-Photon Intravital Microscopy of Glioblastoma in a Murine Model

The delivery of intravenously administered cancer therapeutics to brain tumors is limited by the blood-brain barrier. A method to directly image the accumulation and distribution of macromolecules in brain tumors in vivo would greatly enhance our ability to understand and optimize drug delivery in preclinical models. This protocol describes a method for real-time in vivo tracking of intravenously administered fluorescent-labeled nanoparticles with two-photon intravital microscopy (2P-IVM) in a mouse model of glioblastoma (GBM).
The protocol contains a multi-step description of the procedure, including anesthesia and analgesia of experimental animals, creating a cranial window, GBM cell implantation, placing a head bar, conducting 2P-IVM studies, and post-surgical care for long-term follow-up studies. We show representative 2P-IVM imaging sessions and image analysis, examine the advantages and disadvantages of this technology, and discuss potential applications.
This method can be easily modified and adapted for different research questions in the field of in vivo preclinical brain imaging.

저자 스포트라이트: 교모세포종 치료에서 약물 전달 및 조기 발견 최적화를 위한 멀티모달 이미징 전략

We present a novel approach for two-photon microscopy of the tumor delivery of fluorescent-labeled iron oxide nanoparticles to glioblastoma in a mouse model.

쥐 모델에서 교모세포종의 Two-Photon Intravital Microscopy

The delivery of intravenously administered cancer therapeutics to brain tumors is limited by the blood-brain barrier. A method to directly image the ...

<meta charset="utf8"></head><body>쥐 교모세포종에서 영상 유도 치료 전달을 위한 산화철 나노입자

Nanoparticle Delivery of an Oligonucleotide Payload in a Glioblastoma Multiforme Animal Model

Glioblastoma multiforme (GBM) is the most common and aggressive form of primary brain malignancy for which there is no cure. The blood-brain barrier is a significant hurdle in the delivery of therapies to GBM. Reported here is an image-guided, iron oxide-based therapeutic delivery nano platform capable of bypassing this physiological barrier by virtue of size and accumulating in the tumor region, delivering its payload. This 25 nm nano platform consists of crosslinked dextran-coated iron oxide nanoparticles labeled with Cy5.5 fluorescent dye and containing antisense oligonucleotide as a payload. The magnetic iron oxide core enables tracking of the nanoparticles through in vivo magnetic resonance imaging, while Cy5.5 dye allows tracking by optical imaging. This report details the monitoring of the accumulation of this nanoparticle platform (termed MN-anti-miR10b) in orthotopically implanted glioblastoma tumors following intravenous injection. In addition, it provides insight into the in vivo delivery of RNA oligonucleotides, a problem that has hampered the translation of RNA therapeutics into the clinic.

<meta charset="utf8"></head><body>저자 스포트라이트: 교모세포종 치료를 위한 산화철 나노입자를 사용한 혁신적인 암 치료법

The blood-brain barrier is a significant hurdle in the delivery of therapies for glioblastoma, a disease for which there is no cure. Here, we report an in vivo image-guided iron oxide therapeutic nano platform that can bypass this physiological barrier by virtue of size and accumulate in the tumor.

Glioblastoma multiforme animal model에서 oligonucleotide payload의 나노입자 전달

Glioblastoma multiforme (GBM) is the most common and aggressive form of primary brain malignancy for which there is no cure. The blood-brain barrier is a ...

이후 3D로 Intracranial 주입 VIVO에서 발광 생물 이미징

필기록

요약

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이 비디오의 챕터

Bioluminescence 이미징을 사용하여 종양 성장과 치료에 반응 이후 분석 Intracranial 뇌종양의 Xenografts 구축

유방암 뇌 전이의 종양 Hypoxia 역학 생체내의 Bioluminescence 이미징에서는 마우스 모델에서

Vivo에서 이미징 세포종 Xenografts의 형광 분자 단층 촬영

Stereotactic Intracranial 주입 및

소설 고해상도

소성을 Glioblastoma의 마우스 모델은 뇌 실질 물리적 제약 조건을 유지하고의 intravital 두 광자 현미경에 적합

쥐 모델에서 교모세포종의 Two-Photon Intravital Microscopy

쥐 모델에서 교모세포종의 Two-Photon Intravital Microscopy

쥐 모델에서 교모세포종의 Two-Photon Intravital Microscopy

Glioblastoma multiforme animal model에서 oligonucleotide payload의 나노입자 전달