Name: modeling and imaging 3-dimensional collective cell invasion
Uploaded: 2011-12-07T13:00:00
Description: Watch this Scientific Journal Video about Modeling and Imaging 3-Dimensional Collective Cell Invasion at JoVE.com

इस शोध के लिए अनुदान कैंसर रिसर्च यूके से है.

1. फ्लोरोसेंट प्रोटीन के साथ कोशिकाओं के रेट्रोवायरल लेबलिंग <ol><li> प्लेट 0.25 x 10 6 कोशिकाओं पर प्रति अच्छी तरह से एक 6-अच्छी तरह से बाहर रेट्रोवायरल पैकेजिंग कोशिकाओं (जैसे फीनिक्स) 10% भ्रूण गोज...

<ol>
	<li>Olson, M. F., Sahai, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+actin+cytoskeleton+in+cancer+cell+motility.">The actin cytoskeleton in cancer cell motility.</a> Clin. Exp. Metastasis. 26, 273-287 (2008).</li><li>Hoelzinger, D. B., Demuth, T., Berens, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autocrine+factors+that+sustain+glioma+invasion+and+paracrine+biology+in+the+brain+microenvironment.">Autocrine factors that sustain glioma invasion and paracrine biology in the brain microenvironment.</a> J. Natl. Cancer. Inst. 99, 1583-1593 (2007).</li><li>Chaffer, C. L., Weinberg, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+perspective+on+cancer+cell+metastasis.">A perspective on cancer cell metastasis.</a> Science. 331, 1559-1564 (2011).</li><li>Hanahan, D., Weinberg, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Hallmarks+of+Cancer.">The Hallmarks of Cancer.</a> Cell. 100, 57-70 (2000).</li><li>Francia, G., Cruz-Munoz, W., Man, S., Xu, P., Kerbel, R. S. <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+advanced+spontaneous+metastasis+for+experimental+therapeutics.">Mouse models of advanced spontaneous metastasis for experimental therapeutics.</a> Nat. Rev. Cancer. 11, 135-141 (2011).</li><li>Hooper, S., Marshall, J. F., Sahai, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+cell+migration+in+three+dimensions.">Tumor cell migration in three dimensions.</a> Methods. Enzymol. 406, 625-643 (2006).</li><li>Croft, D. R., Olson, 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=Regulating+the+conversion+between+rounded+and+elongated+modes+of+cancer+cell+movement.">Regulating the conversion between rounded and elongated modes of cancer cell movement.</a> Cancer Cell. 14, 349-351 (2008).</li><li>Wolf, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-step+pericellular+proteolysis+controls+the+transition+from+individual+to+collective+cancer+cell+invasion.">Multi-step pericellular proteolysis controls the transition from individual to collective cancer cell invasion.</a> Nat. Cell. Biol. 9, 893-904 (2007).</li><li>Hennigan, R. F., Hawker, K. L., Ozanne, B. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fos-transformation+activates+genes+associated+with+invasion.">Fos-transformation activates genes associated with invasion.</a> Oncogene. 9, 3591-3600 (1994).</li><li>Scott, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=LIM+kinases+are+required+for+invasive+path+generation+by+tumor+and+tumor-associated+stromal+cells.">LIM kinases are required for invasive path generation by tumor and tumor-associated stromal cells.</a> J. Cell. Biol. 191, 169-185 (2010).</li></ol>

Matrigel आक्रमण assays के परंपरागत chemoattractant प्रेरित गतिशीलता के साथ बाह्य मैट्रिक्स प्रोटीन की एक दिशा में परत पर रखा और नीचे में एक फिल्टर के माध्यम से कोशिकाओं के साथ किया गया है स्थापित. Invasiveness कितने कोशिकाओं को फ...

<table border="1"><tbody><tr><th> अभिकर्मक के नाम </th><th> कंपनी </th><th> सूचीपत्र संख्या </th></tr><tr><td> DMEM (Dulbecco संशोधित ईगल मध्यम) </td><td> GIBCO </td><td> 21969 </td></tr><tr><td> भ्रूण गोजातीय सीरम </td><td> Paa </td><td> A15 - 101 </td></tr><tr><td> पेनिसिलीन स्ट्रेप्टोमाइसिन </td><td> GIBCO </td><td> 15140 </td></tr><tr><td> 200 मिमी एल Glutamine (100x) </td><td> GIBCO </td><td> 25050-032 </td></tr><tr><td> Puromycin </td><td> सिग्मा Aldrich </td><td> P8833 </td></tr><tr><td> 0.05% trypsin EDTA </td><td> GIBCO </td><td> 25300 </td></tr><tr><td> Polybrene </td><td> सिग्मा Aldrich </td><td> AL-118 </td></tr><tr><td> Lipofectamine 2000 अभिकर्मक </td><td> Invitrogen </td><td> 11668019 </td></tr><tr><td> 6.5mm Transwells, 8.0 सुक्ष्ममापी ताकना आकार </td><td> कोर्निंग </td><td> 3422 </td></tr><tr><td> पूरा Matrigel </td><td> बी.डी. बायोसाइंसेज </td><td> 354234 </td></tr> &lt;tr&gt; <td> Calcein AM </td><td> Invitrogen </td><td> C1430 </td></tr><tr><td> RNase </td><td> क्विएज़न </td><td> 19101 </td></tr><tr><td> Propidium आयोडाइड </td><td> सिग्मा Aldrich </td><td> P4864 </td></tr><tr><td> Confocal microcope </td><td> Leica </td><td> SP2MP </td></tr></tbody></table>

modeling and imaging 3-dimensional collective cell invasion

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy tissue is the root cause of disease and death. For other cancers (e.g. breast, lung, etc.), it is the process of metastasis, in which tumor cells move from a primary tumor mass, colonize distal sites and ultimately contribute to organ failure, that eventually leads to morbidity and mortality 3. It has been estimated that invasion and metastasis are responsible for 90&#37; of cancer deaths 4. As a result, there has been intense interest in identifying the molecular processes and critical protein mediators of invasion and metastasis for the purposes of improving diagnosis and treatment 5.
A challenge for cancer scientists is to develop invasion assays that sufficiently resemble the in vivo situation to enable accurate disease modeling 6. Two-dimensional cell motility assays are only informative about one aspect of invasion and do not take into account extracellular matrix (ECM) protein remodeling which is also a critical element. Recently, research has refined our understanding of tumor cell invasion and revealed that individual cells may move by elongated or rounded modes 7. In addition, there has been greater appreciation of the contribution of collective invasion, in which cells invade in strands, sheets and clusters, particularly in highly differentiated tumors that maintain epithelial characteristics, to the spread of cancer 8.
We present a refined method 9 for examining the contributions of candidate proteins to collective invasion 10. In particular, by engineering separate pools of cells to express different fluorescent proteins, it is possible to molecularly dissect the activities and proteins required in leading cells versus those required in following cells. The use of RNAi provides the molecular tool to experimentally disassemble the processes involved in individual cell invasion as well as in different positions of collective invasion. In this procedure, mixtures of fluorescently-labeled cells are plated on the bottom of a Transwell insert previously filled with Matrigel ECM protein, then allowed to invade "upwards" through the filter and into the Matrigel. Reconstruction of z-series image stacks, obtained by confocal imaging, into three-dimensional representations allows for visualization of collectively invading strands and analysis of the representation of fluorescently-labeled cells in leading versus following positions.

Watch this Scientific Journal Video about Modeling and Imaging 3-Dimensional Collective Cell Invasion at JoVE.com

Modeling and Imaging 3-Dimensional Collective Cell Invasion

तीन आयामी मैट्रिक्स कोशिकी में ट्यूमर सेल आक्रमण के मॉडल बेहतर प्रतिबिंबित<em&gt; Vivo में</em&gt; दो आयामी गतिशीलता assays की तुलना में स्थिति है. मैट्रिक्स आक्रमण fluorescently लेबल कोशिकाओं आक्रमण मोड और प्रमुख बनाम निम्नलिखित कोशिकाओं के विशिष्ट योगदान पर विस्तृत जानकारी के confocal इमेजिंग के साथ संयुक्त assays का उपयोग प्राप्त किया जा सकता है.

मॉडलिंग और 3 - आयामी इमेजिंग कलेक्टिव सेल आक्रमण

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy ...

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Medicine

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

मौखिक स्क्वैमस सेल कार्सिनोमा के एक Orthotopic माउस मॉडल में ट्यूमर सेल आक्रमण की मल्टी फोटॉन इमेजिंग

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

Spheroid Assay to Measure TGF-&#946;-induced Invasion

Spheroid Assay to Measure TGF-ÃƒÅ½Ã‚Â²-induced Invasion

उपगोल TGF-β प्रेरित आक्रमण उपाय परख

TGF-&beta; has opposing roles in breast cancer progression by acting as a tumor suppressor in the initial phase, but stimulating invasion and metastasis ...

3-Dimensional Resin Casting and Imaging of Mouse Portal Vein or Intrahepatic Bile Duct System

3 आयामी राल कास्टिंग और माउस पोर्टल नस या intrahepatic पित्त नली प्रणाली की इमेजिंग

In  organs, the correct architecture of vascular and ductal structures is  indispensable for proper physiological function, and the formation and  ...

Molecular Imaging to Target Transplanted Muscle Progenitor Cells

आण्विक प्रतिरोपित स्नायु पूर्वपुस्र्ष कोशिकाओं को लक्षित इमेजिंग

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle ...

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

जनरेशन और चूहे में धमनी घावों की 3-आयामी Quantitation ऑप्टिकल प्रक्षेपण टोमोग्राफी का प्रयोग

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

आण्विक कोलोरेक्टल कैंसर की कोशिकाओं का एक 3 आयामी मॉडल में आक्रामक ट्यूमर microenvironment की रूपरेखा और<em&gt; पूर्व vivo</em&gt; Fibroblasts

Invading colorectal cancer (CRC) cells have acquired the capacity to break free from their sister cells, infiltrate the stroma, and remodel the ...

A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

एक संशोधित<em&gt; इन विट्रो</em&gt; आक्रमण परख हार्मोन, मध्यस्थता कैंसर सेल आक्रमण में साइटोकिन्स और / या वृद्धि कारकों की संभावित भूमिका निर्धारित करने के लिए

Blood serum serves as a chemoattractant towards which cancer cells migrate and invade, facilitating their intravasation into microvessels. However, the ...

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

चूहे में सहज metastatic वृक्क कोशिका कार्सिनोमा (mRCC) मॉडलिंग के बाद nephrectomy

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

Isolation and Immortalization of Patient-derived Cell Lines from Muscle Biopsy for Disease Modeling

रोग मॉडलिंग के लिए पेशी बायोप्सी से मरीज व्युत्पन्न सेल लाइनों के अलगाव और अमरता

The generation of patient-specific cell lines represents an invaluable tool for diagnostic or translational research, and these cells can be collected ...

A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting

कैंसर कोशिका उपगोल परख एक 3 डी सेटिंग में आक्रमण का आकलन करने के लिए

The invasive nature of cancer cell lines is thought to correlate with their metastatic potential.  Most traditional assays, however, do not examine these ...