Name: multimodal imaging of stem cell implantation in the central nervous system of mice
Uploaded: 2012-06-13T13:00:00
Description: Watch this Scientific Journal Video about Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice at JoVE.com

<p class="jove_content"&gt; लेखकों काम अनुसंधान अनुदान आईडी BOF 2006 एंटवर्प विश्वविद्यालय (पीपीओ और AVdL तक दी गई), को अनुसंधान G.0136.11 और G.0130.11 अनुदान द्वारा (करने के लिए प्रदान AVdL, ZB और पीपीओ) और 1.5.021.09 द्वारा समर्थित किया गया था. N.00 के लिए कोष में फ्लेमी विज्ञान और प्रौद्योगिकी के लिए संस्थान की BRAINSTIM (ZB और AVDL दी गई): वैज्ञानिक अनुसंधान फ़्लैंडर्स (FWO-Vlaanderen, बेल्जियम), SBO अनुसंधान आईडब्ल्यूटी 60,838 अनुदान द्वारा (पीपीओ करने के लिए दी गई) Flemish सरकार की ओर से एक Methusalem अनुसंधान अनुदान (ZB के लिए दी गई), चुनाव आयोग FP6-Noe Dimi (LSHB CT-2005-+५१२१४६) द्वारा भाग में, चुनाव आयोग FP6-Noe एमिल (LSHC CT-2004-५०३५६९) द्वारा भाग , और अंतर विश्वविद्यालय आकर्षण डंडे में से IUAP-NIMI-P6/38 (AVDL के लिए दी गई). नथाली डी Vocht एक व्लान्देरेन FWO - से पीएचडी - छात्रवृत्ति रखती है. पीटर Ponsaerts व्लान्देरेन FWO - एक के बाद डॉक्टरेट साथी है.</p

1. सेल तैयार <ol><li> प्रयोगों पूर्व vivo सुसंस्कृत स्टेम सेल आनुवंशिक luciferase और EGFP संवाददाता प्रोटीन व्यक्त इंजीनियर आबादी का उपयोग शुरू किया जाना चाहिए. यहाँ हम / luciferase EGFP व्यक्त murine व्युत्पन्न अस्थि मज्जा stromal कोशि?...

<ol>
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R., Holzapfel, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carboxylated+superparamagnetic+iron+oxide+particles+label+cells+intracellularly+without+transfection+agents.">Carboxylated superparamagnetic iron oxide particles label cells intracellularly without transfection agents.</a> Mol Imaging Biol. 10, 138-146 (2008).</li><li>Modo, M., Hoehn, M., Bulte, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+MR+imaging.">Cellular MR imaging.</a> Mol. Imaging. 4, 143-164 (2005).</li><li>Hinds, K. A., Hill, J. M., Shapiro, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+efficient+endosomal+labeling+of+progenitor+and+stem+cells+with+large+magnetic+particles+allows+magnetic+resonance+imaging+of+single+cells.">Highly efficient endosomal labeling of progenitor and stem cells with large magnetic particles allows magnetic resonance imaging of single cells.</a> Blood. 102, 867-872 (2003).</li><li>Shapiro, E. M., Sharer, K., Skrtic, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+detection+of+single+cells+by+MRI.">In vivo detection of single cells by MRI.</a> Magn. Reson. Med. 55, 242-249 (2006).</li><li>Shapiro, E. M., Skrtic, S., Sharer, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI+detection+of+single+particles+for+cellular+imaging.">MRI detection of single particles for cellular imaging.</a> Proc. Natl. Acad. Sci. U.S.A. 101, 10901-10906 (2004).</li><li>Crabbe, A., Vandeputte, C., Dresselaers, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+MRI+contrast+agents+on+the+stem+cell+phenotype.">Effects of MRI contrast agents on the stem cell phenotype.</a> Cell Transplant. 19, 919-936 (2010).</li><li>Szarecka, A., Xu, Y., Tang, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamics+of+firefly+luciferase+inhibition+by+general+anesthetics:+Gaussian+and+anisotropic+network+analyses.">Dynamics of firefly luciferase inhibition by general anesthetics: Gaussian and anisotropic network analyses.</a> Biophys. J. 93, 1895-1905 (2007).</li><li>Keyaerts, M., Heneweer, C., Gainkam, L. 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इस रिपोर्ट में, हम CNS में प्रतिरक्षा सक्षम चूहों के सेलुलर प्रत्यारोपण की विस्तृत लक्षण वर्णन के लिए तीन पूरक इमेजिंग रूपात्मकता के संयोजन (BLI, एमआरआई और ऊतक विज्ञान) के लिए एक अनुकूलित प्रोटोकॉल का वर्ण...

<table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td><td> टिप्पणियाँ </td></tr><tr><td> IMDM </td><td> LONZA </td><td> BE12-722F </td><td> सेल के विकास मध्यम CEM के घटक </td></tr><tr><td> भ्रूण गोजातीय सीरम </td><td> Gibco </td><td> 10270-106 </td><td> सेल के विकास मध्यम CEM के घटक </td></tr><tr><td> हार्स सीरम </td><td> Gibco </td><td> 1605-122 </td><td> सेल के विकास मध्यम CEM के घटक </td></tr><tr><td> पेनिसिलिन - स्ट्रेप्टोमाइसिन </td><td> Gibco </td><td> 15,140 </td><td> सेल के विकास मध्यम CEM के घटक </td></tr><tr><td> Fungizone </td><td> Gibco </td><td> 15290-018 </td><td> सेल के विकास मध्यम CEM के घटक </td></tr><tr><td> पीबीएस </td><td> Gibco </td><td> 14,190 </td><td></td></tr><tr><td> Puromycine </td><td> Invivogen </td><td> चींटी-जनसंपर्क 1 </td><td></td></tr><tr><td> क्लोम - रस </tघ&gt; <td> Gibco </td><td> +२५३०० </td><td></td></tr><tr><td> जीबी MPIO </td><td> बैंग्स प्रयोगशालाओं </td><td> ME04F/7833 </td><td></td></tr><tr><td> D-luciferin </td><td> Promega </td><td> E1601 </td><td></td></tr><tr><td> Ketamine (Ketalar) </td><td> फ़िज़र </td><td></td><td></td></tr><tr><td> Xylazine (Rompun) </td><td> बायर स्वास्थ्य देखभाल </td><td></td><td></td></tr><tr><td> Isoflurane </td><td> Isoflo </td><td> 05260-05 </td><td></td></tr><tr><td> 0.9% NaCl समाधान </td><td> बैक्सटर </td><td></td><td></td></tr><tr><td> paraformaldehyde </td><td> मर्क </td><td> 1.04005.1000 </td><td></td></tr><tr><td> सूक्रोज </td><td> Applichem </td><td> A1125 </td><td></td></tr><tr><td> माइक्रो इंजेक्शन पंप </td><td> केडी वैज्ञानिक </td><td> KDS100 </td><td></td></tr><tr><td> फोटॉन imager </td><td> Biospace लैब </td><td></td><td></td></tr><tr&gt; <td> 9.4T एमआर स्कैनर </td><td> Bruker BioSpin </td><td> 94 biospec / 20 usr </td><td></td></tr><tr><td> BX51 खुर्दबीन </td><td> ओलिंप </td><td> BX51 </td><td></td></tr><tr><td> Mycrom एचएम cryostat </td><td> Prosan </td><td> HM525 </td><td></td></tr><tr><td> सिरिंज </td><td> हैमिलटन </td><td> 7635-01 </td><td></td></tr><tr><td> 30 गेज सुई </td><td> हैमिलटन </td><td> 7762-03 </td><td></td></tr><tr><td> फोटो विजन सॉफ्टवेयर </td><td> Biospace लैब </td><td></td><td></td></tr><tr><td> M3vision सॉफ्टवेयर </td><td> Biospace लैब </td><td></td><td></td></tr><tr><td> Paravision 5.1 सॉफ्टवेयर </td><td> Bruker BioSpin </td><td></td><td></td></tr><tr><td> अमीरा 4.0 सॉफ्टवेयर </td><td> चेहरा इमेजिंग </td><td></td><td></td></tr></tbody></table>

multimodal imaging of stem cell implantation in the central nervous system of mice

During the past decade, stem cell transplantation has gained increasing interest as primary or secondary therapeutic modality for a variety of diseases, both in preclinical and clinical studies. However, to date results regarding functional outcome and/or tissue regeneration following stem cell transplantation are quite diverse. Generally, a clinical benefit is observed without profound understanding of the underlying mechanism(s)1. Therefore, multiple efforts have led to the development of different molecular imaging modalities to monitor stem cell grafting with the ultimate aim to accurately evaluate survival, fate and physiology of grafted stem cells and/or their micro-environment. Changes observed in one or more parameters determined by molecular imaging might be related to the observed clinical effect. In this context, our studies focus on the combined use of bioluminescence imaging (BLI), magnetic resonance imaging (MRI) and histological analysis to evaluate stem cell grafting.
BLI is commonly used to non-invasively perform cell tracking and monitor cell survival in time following transplantation2-7, based on a biochemical reaction where cells expressing the Luciferase-reporter gene are able to emit light following interaction with its substrate (e.g. D-luciferin)8, 9. MRI on the other hand is a non-invasive technique which is clinically applicable10 and can be used to precisely locate cellular grafts with very high resolution11-15, although its sensitivity highly depends on the contrast generated after cell labeling with an MRI contrast agent. Finally, post-mortem histological analysis is the method of choice to validate research results obtained with non-invasive techniques with highest resolution and sensitivity. Moreover end-point histological analysis allows us to perform detailed phenotypic analysis of grafted cells and/or the surrounding tissue, based on the use of fluorescent reporter proteins and/or direct cell labeling with specific antibodies.
In summary, we here visually demonstrate the complementarities of BLI, MRI and histology to unravel different stem cell- and/or environment-associated characteristics following stem cell grafting in the CNS of mice. As an example, bone marrow-derived stromal cells, genetically engineered to express the enhanced Green Fluorescent Protein (eGFP) and firefly Luciferase (fLuc), and labeled with blue fluorescent micron-sized iron oxide particles (MPIOs), will be grafted in the CNS of immune-competent mice and outcome will be monitored by BLI, MRI and histology (Figure 1).

Watch this Scientific Journal Video about Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice at JoVE.com

Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice

यह लेख कृंतक मस्तिष्क में सेलुलर grafts के का उपयोग करने के बहुविध इमेजिंग के लिए घटनाओं की एक अनुकूलित अनुक्रम का वर्णन: (i) vivo में bioluminescence और चुंबकीय अनुनाद इमेजिंग में, और (ii) पोस्टमार्टम ऊतकीय विश्लेषण. एक ही जानवर पर इन इमेजिंग रूपात्मकता के संयोजन उच्च संकल्प, संवेदनशीलता और विशिष्टता के साथ सेलुलर भ्रष्टाचार के मूल्यांकन की अनुमति देता है.

चूहे के केंद्रीय तंत्रिका तंत्र में स्टेम सेल आरोपण के मल्टीमॉडल इमेजिंग

During the past decade, stem cell transplantation has gained increasing interest as primary or secondary therapeutic modality for a variety of diseases, ...

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Neuroscience

Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System

ऑप्टिक तंत्रिका transection: केन्द्रीय तंत्रिका तंत्र में वयस्क न्यूरॉन Apoptosis के एक मॉडल

Retinal ganglion cells (RGCs) are CNS neurons that output visual information from the retina to the brain, via the optic nerve. 
The optic nerve can be ...

Production of Lentiviral Vectors for Transducing Cells from the Central Nervous System

केन्द्रीय तंत्रिका तंत्र से Transducing कक्ष के लिए lentiviral वैक्टर का उत्पादन

Efficient gene delivery in the central nervous system (CNS) is important in studying gene functions, modeling neurological diseases and developing ...

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

केन्द्रीय तंत्रिका तंत्र में एकल कक्षों के लेबल<em&gt; ड्रोसोफिला मेलानोगास्टर</em

In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

Direct Intraventricular Delivery of Drugs to the Rodent Central Nervous System

कृंतक केन्द्रीय तंत्रिका तंत्र के लिए ड्रग्स का प्रत्यक्ष intraventricular डिलिवरी

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses ...

In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation

In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation

Polyribosome Fractionation द्वारा केंद्रीय तंत्रिका तंत्र Translatome की विवो पूछताछ में

Multiple processes are involved in gene expression including transcription, translation and stability of mRNAs and proteins. Each of these steps are ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

कृंतक केंद्रीय तंत्रिका तंत्र के विवो optogenetic उत्तेजना में

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution ...

Functional Evaluation of Biological Neurotoxins in Networked Cultures of Stem Cell-derived Central Nervous System Neurons

स्टेम नेटवर्क संस्कृतियों में जैविक neurotoxins के कार्यात्मक मूल्यांकन केन्द्रीय तंत्रिका तंत्र न्यूरॉन्स सेल व्युत्पन्न

Therapeutic and mechanistic studies of the presynaptically targeted clostridial neurotoxins (CNTs) have been limited by the need for a scalable, ...

In Situ Ca2+ Imaging of the Enteric Nervous System

In Situ Ca2+ Imaging of the Enteric Nervous System

आंतों का तंत्रिका तंत्र की स्वस्थानी सीए 2 + इमेजिंग में

Reflex behaviors of the intestine are controlled by the enteric nervous system (ENS). The ENS is an integrative network of neurons and glia in two ...

Optical Clearing of the Mouse Central Nervous System Using Passive CLARITY

माउस केंद्रीय तंत्रिका तंत्र के ऑप्टिकल क्लियरिंग निष्क्रिय स्पष्टता का उपयोग

Traditionally, tissue visualization has required that the tissue of interest be serially sectioned and imaged, subjecting each tissue section to unique ...

Electrophysiological Recording of The Central Nervous System Activity of Third-Instar Drosophila Melanogaster

Electrophysiological Recording of The Central Nervous System Activity of Third-Instar  Drosophila Melanogaster

तीसरी-Instar Drosophila Melanogaster के केंद्रीय तंत्रिका तंत्र गतिविधि की Electrophysiological रिकॉर्डिंग

The majority of the currently available insecticides target the nervous system and genetic mutations of invertebrate neural proteins oftentimes yield ...