Name: generation of microtumors using 3d human biogel culture system and patient-derived glioblastoma cells for kinomic profiling and drug response testing
Uploaded: 2016-06-09T15:00:00
Description: Watch this Scientific Journal Video about Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing at JoVE.

, ब्रेन ट्यूमर बीजाणु पुरस्कार (पीडी: GY गिलेस्पी, P20CA 151129-03) और एसबीआईआर अनुबंध (पीआई: आर सिंह, N43CO-2013-00026): एनआईएच R21 अनुदान (सी विले, CA185712-01 पीआई) द्वारा समर्थित है।

नोट: सभी जेनोग्राफ्ट चिकित्सा मूल्यांकन एक प्रोटोकॉल संस्थागत पशु की देखभाल और उपयोग समिति द्वारा अनुमोदित पर ग्लियोब्लास्टोमा के लिए एक orthotopic ट्यूमर मॉडल का उपयोग किया गया। 1. रोगी व्युत्प?...

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	<li>Ohgaki, H., Kleihues, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Population-based+studies+on+incidence,+survival+rates,+and+genetic+alterations+in+astrocytic+and+oligodendroglial+gliomas.">Population-based studies on incidence, survival rates, and genetic alterations in astrocytic and oligodendroglial gliomas.</a> J Neuropathol Exp Neurol. 64 (6), 479-489 (2005).</li><li>Wen, P. Y., Kesari, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Malignant+gliomas+in+adults.">Malignant gliomas in adults.</a> N Engl J Med. 359 (5), 492-507 (2008).</li><li>Thumma, S. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+pretreatment+clinical+factors+on+overall+survival+in+glioblastoma+multiforme:+a+Surveillance+Epidemiology+and+End+Results+(SEER)+population+analysis.">Effect of pretreatment clinical factors on overall survival in glioblastoma multiforme: a Surveillance Epidemiology and End Results (SEER) population analysis.</a> World J Surg Oncol. 10 (75), (2012).</li><li>Furnari, F. B., Cloughesy, T. F., Cavenee, W. K., Mischel, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneity+of+epidermal+growth+factor+receptor+signalling+networks+in+glioblastoma.">Heterogeneity of epidermal growth factor receptor signalling networks in glioblastoma.</a> Nat Rev Cancer. 15 (5), 302-310 (2015).</li><li>Mischel, P. S., Cloughesy, T. F., Nelson, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA-microarray+analysis+of+brain+cancer:+molecular+classification+for+therapy.">DNA-microarray analysis of brain cancer: molecular classification for therapy.</a> Nat Rev Neurosci. 5 (10), 782-792 (2004).</li><li>Verhaak, R. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrated+genomic+analysis+identifies+clinically+relevant+subtypes+of+glioblastoma+characterized+by+abnormalities+in+PDGFRA,+IDH1,+EGFR,+and+NF1.">Integrated genomic analysis identifies clinically relevant subtypes of glioblastoma characterized by abnormalities in PDGFRA, IDH1, EGFR, and NF1.</a> Cancer Cell. 17 (1), 98-110 (2010).</li><li>De Witt Hamer, P. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genomic+profile+of+human+malignant+glioma+is+altered+early+in+primary+cell+culture+and+preserved+in+spheroids.">The genomic profile of human malignant glioma is altered early in primary cell culture and preserved in spheroids.</a> Oncogene. 27 (14), 2091-2096 (2008).</li><li>Shankavaram, U. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+profiling+indicates+orthotopic+xenograft+of+glioma+cell+lines+simulate+a+subclass+of+human+glioblastoma.">Molecular profiling indicates orthotopic xenograft of glioma cell lines simulate a subclass of human glioblastoma.</a> J Cell Mol Med. 16 (3), 545-554 (2012).</li><li>Abbott, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+culture:+biology's+new+dimension.">Cell culture: biology's new dimension.</a> Nature. 424 (6951), 870-872 (2003).</li><li>Rao, S. S., Lannutti, J. J., Viapiano, M. S., Sarkar, A., Winter, J. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+3D+biomimetic+models+to+understand+the+behavior+of+glioblastoma+multiforme+cells.">Toward 3D biomimetic models to understand the behavior of glioblastoma multiforme cells.</a> Tissue Eng Part B Rev. 20 (4), 314-327 (2014).</li><li>Pampaloni, F., Reynaud, E. G., Stelzer, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+third+dimension+bridges+the+gap+between+cell+culture+and+live+tissue.">The third dimension bridges the gap between cell culture and live tissue.</a> Nat Rev Mol Cell Biol. 8 (10), 839-845 (2007).</li><li>Hollister, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Porous+scaffold+design+for+tissue+engineering.">Porous scaffold design for tissue engineering.</a> Nat Mater. 4 (7), 518-524 (2005).</li><li>Rowley, J. A., Madlambayan, G., Mooney, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alginate+hydrogels+as+synthetic+extracellular+matrix+materials.">Alginate hydrogels as synthetic extracellular matrix materials.</a> Biomaterials. 20 (1), 45-53 (1999).</li><li>Hughes, C. S., Postovit, L. M., Lajoie, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrigel:+a+complex+protein+mixture+required+for+optimal+growth+of+cell+culture.">Matrigel: a complex protein mixture required for optimal growth of cell culture.</a> Proteomics. 10 (9), 1886-1890 (2010).</li><li>Yoshino, J. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proliferation+and+differentiation+of+a+transfected+Schwann+cell+line+is+altered+by+an+artificial+basement+membrane.">Proliferation and differentiation of a transfected Schwann cell line is altered by an artificial basement membrane.</a> Glia. 3 (5), 315-321 (1990).</li><li>Siegal, G. P., Singh, R., Foundation, T. U. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biologically+active+native+biomatrix+composition.">Biologically active native biomatrix composition.</a> US patent. , (2010).</li><li>Sarkaria, J. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+an+orthotopic+xenograft+model+for+assessing+the+effect+of+epidermal+growth+factor+receptor+amplification+on+glioblastoma+radiation+response.">Use of an orthotopic xenograft model for assessing the effect of epidermal growth factor receptor amplification on glioblastoma radiation response.</a> Clin Cancer Res. 12 (7), 2264-2271 (2006).</li><li>Strober, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trypan+Blue+Exclusion+Test+of+Cell+Viability.">Trypan Blue Exclusion Test of Cell Viability.</a> Curr Protoc Immunol. 111, 1-3 (2015).</li><li>Anderson, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinomic+exploration+of+temozolomide+and+radiation+resistance+in+Glioblastoma+multiforme+xenolines.">Kinomic exploration of temozolomide and radiation resistance in Glioblastoma multiforme xenolines.</a> Radiother Oncol. 111 (3), 468-474 (2014).</li><li>Anderson, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinomic+profiling+of+electromagnetic+navigational+bronchoscopy+specimens:+a+new+approach+for+personalized+medicine.">Kinomic profiling of electromagnetic navigational bronchoscopy specimens: a new approach for personalized medicine.</a> PLoS One. 9 (12), 116388 (2014).</li><li>Jarboe, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinomic+profiling+approach+identifies+Trk+as+a+novel+radiation+modulator.">Kinomic profiling approach identifies Trk as a novel radiation modulator.</a> Radiother Oncol. 103 (3), 380-387 (2012).</li><li>Anderson, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+Throughput+Kinomic+Profiling+of+Human+Clear+Cell+Renal+Cell+Carcinoma+Identifies+Kinase+Activity+Dependent+Molecular+Subtypes.">High Throughput Kinomic Profiling of Human Clear Cell Renal Cell Carcinoma Identifies Kinase Activity Dependent Molecular Subtypes.</a> PLoS One. 10 (9), 0139267 (2015).</li><li>Anderson, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinomic+Alterations+in+Atypical+Meningioma.">Kinomic Alterations in Atypical Meningioma.</a> Medical Research Archives. 3, (2015).</li><li>Hothi, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+chemical+screens+identify+disulfiram+as+an+inhibitor+of+human+glioblastoma+stem+cells.">High-throughput chemical screens identify disulfiram as an inhibitor of human glioblastoma stem cells.</a> Oncotarget. 3 (10), 1124-1136 (2012).</li><li>Quartararo, C. E., Reznik, E., deCarvalho, A. C., Mikkelsen, T., Stockwell, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-Throughput+Screening+of+Patient-Derived+Cultures+Reveals+Potential+for+Precision+Medicine+in+Glioblastoma.">High-Throughput Screening of Patient-Derived Cultures Reveals Potential for Precision Medicine in Glioblastoma.</a> ACS Med Chem Lett. 6 (8), 948-952 (2015).</li><li>Ma, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Towards+personalized+medicine+with+a+three-dimensional+micro-scale+perfusion-based+two-chamber+tissue+model+system.">Towards personalized medicine with a three-dimensional micro-scale perfusion-based two-chamber tissue model system.</a> Biomaterials. 33 (17), 4353-4361 (2012).</li><li>Pedron, S., Becka, E., Harley, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+glioma+cell+phenotype+in+3D+matrices+by+hyaluronic+acid.">Regulation of glioma cell phenotype in 3D matrices by hyaluronic acid.</a> Biomaterials. 34 (30), 7408-7417 (2013).</li><li>Rape, A., Ananthanarayanan, B., Kumar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+strategies+to+mimic+the+glioblastoma+microenvironment.">Engineering strategies to mimic the glioblastoma microenvironment.</a> Adv Drug Deliv Rev. 79-90, 172-183 (2014).</li><li>Willey, C. D., Gilbert, A. N., Anderson, J. C., Gillespie, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patient-Derived+Xenografts+as+a+Model+System+for+Radiation+Research.">Patient-Derived Xenografts as a Model System for Radiation Research.</a> Semin Radiat Oncol. 25 (4), 273-280 (2015).</li></ol>

प्रोटोकॉल के भीतर महत्वपूर्ण कदम मुख्य रूप से पीढ़ी है, साथ ही दवा खुराक और रखरखाव microtumor से संबंधित हैं। क्योंकि microtumor मोती नाजुक और आसानी से फाड़ रहे हैं, अत्यधिक ध्यान दोनों एक परख और रखरखाव के विकास के च?...

सबसे आम प्राथमिक intracranial घातक ब्रेन ट्यूमर ग्रेड III astrocytomas और ग्रेड चतुर्थ ग्लियोब्लास्टोमा मल्टीफार्मी (ग्लियोब्लास्टोमा या जीबीएम) कर रहे हैं। अमेरिका में 1-3 जीबीएम के लिए वर्तमान चिकित्सा के साथ 15 माह - ये ट्यूमर 12 के बीच मंझला एक साल के अस्तित्व के साथ गरीब prognoses प्रदान करते हैं। Multimodality उपचारों सर्जरी, विकिरण, और temozolomide (TMZ) और काइनेज-लक्षित एजेंट सहित कीमोथेरेपी शामिल हैं। काइनेज सिगनल अक्सर जीबीएम में dysregulated है epidermal वृद्धि कारक रिसेप्टर (EGFR), प्लेटलेट व्युत्पन्न वृद्धि कारक रिसेप्टर (PDGFR) में वृद्धि के संकेत, बढ़ Phosphatidyl-Inositol -3 kinase (PI3K) और प्रवर्धन के साथ ट्यूमर के सबसेट या सक्रिय म्यूटेशन सहित, ट्यूमर संवहनी endothelial वृद्धि कारक रिसेप्टर (VEGFR) के साथ ही अन्य काइनेज संचालित रास्ते 4-6 के माध्यम से एन्जियोजेनिक संकेतन का समर्थन। इन विट्रो में और vivo मॉडल अक्सर ये प्रतिनिधि परिवर्तन खो वर्तमान &lt;sup&gt; 7। इसके अतिरिक्त, आनुवंशिक प्रोफाइलिंग प्रत्याशित लाभ है कि तथ्य यह है कि आनुवंशिक और epigenetic परिवर्तन हमेशा प्रोटीन गतिविधि का स्तर है, जहां सबसे काइनेज को निशाना एजेंटों सीधे अधिनियम में परिवर्तन की भविष्यवाणी नहीं है, और जहां कार्रवाई की अन्य तंत्र के साथ चिकित्सा कार्य कर सकते हैं प्रतिबिंबित कर सकते हैं की पेशकश नहीं किया गया है परोक्ष रूप से। पारंपरिक अमर सेल लाइन है कि विज्ञापन अनन्त passaged किया जा सकता भर के रखरखाव और reproducibility के अपने आसानी के कारण दवा के परीक्षण के लिए मानक किया गया है। हालांकि, इस मॉडल एक उच्च पोषक तत्व (और कृत्रिम) विकास वातावरण है कि तेजी से कोशिकाओं है कि मूल ट्यूमर से काफी भिन्न उगाने के लिए चुनता से ग्रस्त है। जैसे, वहाँ और अधिक यथार्थवादी मॉडल प्रणाली के रूप में रोगी में मौजूद है कि एक अधिक जटिल ट्यूमर जैविक प्रणाली को प्रतिबिंबित विकसित करने में काफी रुचि रही है। ट्यूमर xenografts एक प्राथमिक ट्यूमर चूहों में उगाया ( &quot;xenoline,&quot; मरीज व्युत्पन्न जेनोग्राफ्ट या PDX) प्रावधानों से सीधे विकसितडी एक अधिक चिंतनशील मॉडल प्रणाली, विशेष रूप से कैंसर चिकित्सा विज्ञान की स्थापना में, के रूप में वे और अधिक मज़बूती से नैदानिक ​​सफलता की भविष्यवाणी करने के लिए। 8 अधिक चिंतनशील जीव विज्ञान के बावजूद महसूस कर रहे हैं, इन मॉडलों महंगे हैं और स्थापित करने और बनाए रखने के लिए मुश्किल हो जाता है। इसके अलावा, वे उच्च throughput अध्ययन करने के लिए उत्तरदायी नहीं हैं। जरूरत को बेहतर जीवविज्ञान का मॉडल है कि अधिक सही प्राथमिक ट्यूमर में आणविक परिवर्तन को प्रतिबिंबित, और प्रोफ़ाइल के लिए और kinase गतिविधि के प्रत्यक्ष उपायों का उपयोग इन मॉडलों का परीक्षण, आनुवंशिक मार्करों सरोगेट नहीं विकसित करने के लिए, स्पष्ट है। यह अच्छी तरह से मान्यता प्राप्त है कि दो-आयामी (2 डी) monolayer संस्कृतियों के विपरीत, 3 डी या बहुकोशिकीय परख मॉडल और अधिक physiologically प्रासंगिक समापन 9-11 प्रदान कर सकते हैं। आम 3D संस्कृति मैट्रिक्स लेपित microcarriers और सेल अंडाकार आकृति के गठन को शामिल दृष्टिकोण। ट्यूमर spheroids स्पिनर फ्लास्क, pHEMA प्लेट का उपयोग और ड्रॉप तकनीक फांसी सेलुलर एकत्रीकरण के माध्यम से उत्पन्न किया जा सकता है। टी के लिए सीमाओंhese दृष्टिकोण शामिल हैं: कुछ कोशिकाओं स्थिर spheroids, परिवर्तनशीलता विकास में और मिश्रित प्रकार की कोशिकाओं के साथ चुनौतियों के रूप में करने के लिए असमर्थता। वैकल्पिक रूप से, कई सिंथेटिक (हाइड्रोजेल, बहुलक) और पशु व्युत्पन्न Engelbreth-होल्म-झुंड (EHS) माउस सार्कोमा से मैट्रिक्स, गोजातीय कोलेजन) matrices 3 डी संस्कृति के लिए विकसित किया गया है अध्ययन 12-14। माउस EHS मैट्रिक्स बड़े पैमाने पर इस्तेमाल किया लेकिन इन विट्रो और इन विवो 15 में सेल के विकास और भेदभाव को बढ़ावा देने के लिए जाना जाता है। आदेश में 3 डी ट्यूमर जीव विज्ञान को दोहराने के लिए, एक मानव biomatrix प्रणाली डॉ। राज सिंह एट अल 16 से विकसित किया गया था। प्राकृतिक, वृद्धि कारक मुक्त मानव biogel 3D संस्कृति scaffolds (मोती, डिस्क) है, जो अनेक प्रकार की कोशिकाओं के लंबे समय तक खेती का समर्थन करता है। 3 डी मानव biogel संस्कृति डिजाइन की एक श्रृंखला ट्यूमर के विकास, आसंजन, angiogenesis और आक्रमण गुण के अध्ययन के लिए स्थापित कर रहे हैं। फायदे और मानव biogel के गुणों को आम की तुलना मेंमाउस EHS जैल तालिका 1 और 2 टेबल में संक्षेप हैं। <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> स्रोत: </td><td> मानव Amnions (जमा ऊतक) पैथोजन मुक्त, आईआरबी मुक्त / अनुमोदित </td></tr><tr><td> ईसीएम प्रकृति: </td><td> गैर विकृत Biogel (जीएलपी उत्पादन) </td></tr><tr><td> कुंजी अवयव: </td><td> कर्नल-मैं (38%), laminin (22%), कर्नल IV (20%), कर्नल III (7%), Entactin और HSPG (&lt;3%) </td></tr><tr><td> GF-मुक्त: </td><td> Undetectable EGF, FGF, TGF, वीईजीएफ़, PDGF (गैर-एन्जियोजेनिक, गैर विषैले) </td></tr></tbody></table> तालिका 1: मानव Biogel के गुण के रूप में आम EHS जैल की तुलना में। <table border="1" fo:keep-together.within-page="1" fo:keep-wiवें-next.within-पेज = &quot;हमेशा&quot;&gt; <tbody><tr><td> मानव Biogel </td><td> EHS जैल </td></tr><tr><td> प्राकृतिक मानव मैट्रिक्स </td><td> पुनर्गठन माउस मैट्रिक्स </td></tr><tr><td> नियंत्रित सेल के विकास और भेदभाव </td><td> सेल के विकास और भेदभाव को बढ़ावा देने के कर सकते हैं </td></tr><tr><td> शारीरिक जीन अभिव्यक्ति </td><td> चर जीन अभिव्यक्ति </td></tr><tr><td> 3 डी ऊतक की तरह संस्कृति मॉडल </td><td> प्लेट आधारित संस्कृति मॉडल </td></tr></tbody></table> तालिका 2: मानव Biogel के लाभ के रूप में आम EHS जैल की तुलना में।

<table border="0" cellpadding="0" cellspacing="0" width="1142">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Collagenase-I&#160;</td>
			<td style="width:184px;">Sigma-Aldrich</td>
			<td style="width:119px;">CO130</td>
			<td style="width:496px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Trypsin EDTA (10X)</td>
			<td style="width:184px;">Invitrogen</td>
			<td style="width:119px;">15400-054&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Neurobasal-A</td>
			<td style="width:184px;">Life Technologies</td>
			<td style="width:119px;">10888-022</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">N-2 Supplement</td>
			<td style="width:184px;">Life Technologies</td>
			<td style="width:119px;">17502-048</td>
			<td>1x final concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">B-27 Supplement w/o Vitamin A</td>
			<td style="width:184px;">Life Technologies</td>
			<td style="width:119px;">12587-010</td>
			<td>1x final concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Recombinant Human FGF-basic</td>
			<td style="width:184px;">Life Technologies</td>
			<td style="width:119px;">PHG0266</td>
			<td>10 ng/mL final concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Recombinant Human EGF</td>
			<td style="width:184px;">Life Technologies</td>
			<td style="width:119px;">PGH0315</td>
			<td>10 ng/mL final concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">L-Glutamine</td>
			<td style="width:184px;">Corning Cellgro Mediatech</td>
			<td style="width:119px;">25-005-CI</td>
			<td>2 mM final concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Fungizone</td>
			<td style="width:184px;">Omega Scientific</td>
			<td style="width:119px;">FG-70</td>
			<td>2.5 ug/mL final concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Penicillin Streptomycin</td>
			<td style="width:184px;">Omega Scientific</td>
			<td style="width:119px;">PS-20</td>
			<td>100 U/mL Penicillin G, 100 ug/mL Streptomycin final concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Gentamicin</td>
			<td style="width:184px;">Life Technologies</td>
			<td style="width:119px;">15750-060</td>
			<td>50 ng/mL final concentration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">MTT</td>
			<td style="width:184px;">Life Technologies</td>
			<td style="width:119px;">M6494</td>
			<td>prepared to 5 mg/mL in PBS and sterile filtered, 1 mg/mL in well</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">SDS</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">BP166</td>
			<td>for MTT lysis buffer, prepared to 10% in 0.01M HCL, 5% in well</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">HCl</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">A144SI-212</td>
			<td>for MTT lysis buffer, prepared to 0.01M with SDS, 5 mM in well</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Calcein AM</td>
			<td style="width:184px;">Life Technologies</td>
			<td style="width:119px;">C1430</td>
			<td>1 mM in DMSO stock, 2 uM in PBS staining solution, 1 uM in well</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Halt&#8217;s Protein Phosphatase Inhibitor cocktail&#160;</td>
			<td style="width:184px;">Pierce ThermoScientific</td>
			<td style="width:119px;">78420</td>
			<td>1:100 ratio in MPER&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Halt&#39;s Protein Protease Inhibitor&#160;</td>
			<td style="width:184px;">Pierce ThermoScientific</td>
			<td style="width:119px;">87786</td>
			<td>1:100 ratio in MPER</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Mammalian Protein Extraction Reagent (MPER)</td>
			<td style="width:184px;">Pierce ThermoScientific</td>
			<td style="width:119px;">PI78501</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Trypan Blue</td>
			<td style="width:184px;">Pierce ThermoScientific</td>
			<td>15250-061</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">DMSO</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">BP231</td>
			<td>for dissolution of calcein AM &#38; compounds</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Phosphate-Buffered Saline without Ca/Mg</td>
			<td style="width:184px;">Lonza</td>
			<td style="width:119px;">17-517Q</td>
			<td>diluted to 1X with MiliQ ultrapure water and sterile filtered (for cell culture)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Dulbecco&#39;s Phosphate-Buffered Saline with Ca/Mg</td>
			<td style="width:184px;">Corning Cellgro Mediatech</td>
			<td style="width:119px;">20-030-CV</td>
			<td>diluted to 1X with MiliQ ultrapure water (for pre-fixation wash)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">10% Neutral Buffered Formalin</td>
			<td style="width:184px;">Protocol</td>
			<td style="width:119px;">032-060</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Trypan Blue</td>
			<td style="width:184px;">Pierce ThermoScientific</td>
			<td>15250-061</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">High Density Hubiogel</td>
			<td style="width:184px;">Vivo Biosciences</td>
			<td style="width:119px;">HDHG-5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Halt&#39;s Protein Phosphatase Inhibitor</td>
			<td style="width:184px;">Pierce</td>
			<td style="width:119px;">78420</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Halt&#39;s Protein Protease Inhibitor</td>
			<td style="width:184px;">Pierce</td>
			<td style="width:119px;">87786</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Mammalian Protein Extraction Reagent (MPER)</td>
			<td style="width:184px;">Thermo Scientific</td>
			<td style="width:119px;">78501</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Protein Tyrosine Kinase (PTK) Array Profiling chip</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">86312</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">PTK kinase buffer</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">36000</td>
			<td>300 &#181;l 10X PK buffer stock in 2.7 ml dH20, catalog number for PTK reagent kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">ATP</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">36000</td>
			<td>catalog number for PTK reagent kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PY20- FITC-conjugated antibody</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">36000</td>
			<td>catalog number for PTK reagent kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PTK Additive</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">32114</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PTK-MM1 tube (10X BSA)</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">36000</td>
			<td>catalog number for PTK reagent kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Serine/Threonine Kinase (STK) Array Profiling chip</td>
			<td style="width:184px;">PamGene</td>
			<td>87102</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">STK kinase buffer</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">32205</td>
			<td>catalog number for STK reagent kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">STK Primary Antibody Mix (DMAB tube)</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">32205</td>
			<td>catalog number for STK reagent kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">FITC-conjugated Secondary Antibody</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">32203</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">STK-MM1 tube (100X BSA)</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">32205</td>
			<td>catalog number for STK reagent kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">STK Antibody Buffer</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;">32205</td>
			<td>catalog number for STK reagent kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Equipment</td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">#11 Blades, sterile</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">3120030</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">#3 scalpel handles, sterile</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">08-913-5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">100mm glass Petri dishes</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">08-748D</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Semicurved forceps</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">12-460-318</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Trypsinizing flask</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">10-042-12B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Magnetic stirrer</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">14-490-200</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">3/4&#34; stir bar</td>
			<td style="width:184px;">Fisher</td>
			<td>14-512-125</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">B-D cell strainer&#160;</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:119px;">#352340</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">B-D 50ml Centrifuge tube</td>
			<td style="width:184px;">Fisher</td>
			<td>#352098</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">PamStation 12</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">BioNavigator 6.0 kinomic analysis software&#160;</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Evolve Kinase Assay Software</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">UpKin App software (upstream kinase prediction)</td>
			<td style="width:184px;">PamGene</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">gentleMACS Dissociator</td>
			<td style="width:184px;">Miltenyi Biotec</td>
			<td style="width:119px;">130-093-235</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:343px;">Rotary Cell Culture System (RCCS)</td>
			<td style="width:184px;">Synthecon</td>
			<td style="width:119px;">RCCS-D</td>
			<td>with 10 mL disposable HARV</td>
		</tr>
	</tbody>
</table>

generation of microtumors using 3d human biogel culture system and patient-derived glioblastoma cells for kinomic profiling and drug response testing

The use of patient-derived xenografts for modeling cancers has provided important insight into cancer biology and drug responsiveness. However, they are time consuming, expensive, and labor intensive. To overcome these obstacles, many research groups have turned to spheroid cultures of cancer cells. While useful, tumor spheroids or aggregates do not replicate cell-matrix interactions as found in vivo. As such, three-dimensional (3D) culture approaches utilizing an extracellular matrix scaffold provide a more realistic model system for investigation. Starting from subcutaneous or intracranial xenografts, tumor tissue is dissociated into a single cell suspension akin to cancer stem cell neurospheres. These cells are then embedded into a human-derived extracellular matrix, 3D human biogel, to generate a large number of microtumors. Interestingly, microtumors can be cultured for about a month with high viability and can be used for drug response testing using standard cytotoxicity assays such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and live cell imaging using Calcein-AM. Moreover, they can be analyzed via immunohistochemistry or harvested for molecular profiling, such as array-based high-throughput kinomic profiling, which is detailed here as well. 3D microtumors, thus, represent a versatile high-throughput model system that can more closely replicate in vivo tumor biology than traditional approaches.

हम पता चला 3D biogel संस्कृति प्रणाली लंबी अवधि के विकास और अनेक प्रकार की कोशिकाओं के समारोह का समर्थन करता है। इस सहयोगी परियोजना में, रोगी व्युत्पन्न जीबीएम xenolines (PDX) microtumors के सैकड़ों के उत्पादन ?...

Watch this Scientific Journal Video about Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing at JoVE.

Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing

Patient-derived xenografts of glioblastoma multiforme can be miniaturized into living microtumors using 3D human biogel culture system. This in vivo-like 3D tumor assay is suitable for drug response testing and molecular profiling, including kinomic analysis.

Microtumors 3 डी मानव Biogel संस्कृति प्रणाली का प्रयोग और Kinomic रूपरेखा और दवा प्रतिक्रिया परीक्षण के लिए रोगी व्युत्पन्न ग्लयोब्लास्टोमा कोशिकाओं की पीढ़ी

The use of patient-derived xenografts for modeling cancers has provided important insight into cancer biology and drug responsiveness. However, they are ...

The overall goal of this patient derived glioblastoma 3D microtumor culture system is to enable high-throughput drug testing with a corresponding molecular profiling assay, such as kinomic testing. This method can help answer key questions in the cancer field, such as, why do certain therapies work well in one patient's tumor but not in another? The main advantage of this technique is that it provides a miniature yet realistic tumor culture system suitable for therapy testing.Generally, individuals new to this method will struggle because handling the patient derived xenograft cells is not like handling long established, immortalized tumor cells. This is due to their fragility and sensitivity to changing culture conditions. Visual demonstration of this method is critical, as the microtumor generation steps are difficult to learn through description only, due to their unique and complex method of production.Demonstrating the tumor disaggregation procedure will be Catherine Langford, the senior technician and lab manager in my laboratory. Demonstrating the microtumor generation and drug dosing procedures will be Rachael Shevin, a technician in my lab. Using forceps, blunt dissect to gently free the tumor mass from the peritoneal wall and the skin, and transfer the tumor mass to a sterile glass Petri dish.Then, use a sterile scalpel with a number 11 blade and sterile semi-curved forceps to debride the tumor tissue of any necrotic tissue and membranous host connective tissue. Wash the cleaned tumors with sterile RPMI three to five times to remove the excess blood, gently tilting the dish between rinses to remove the wash. After the final wash, use two number 11 scalpel blades to finely mince the tumors.Then, use sterile semi-curved forceps to transfer the tumors into a dissociation tube containing enzyme solution. Place the tube into a cellular dissociator, and select the Tumor 2 program. Click Start"to run the dissociator for 37 seconds.Then transfer the tube to a rotator and place the rotator in a 37 degree Celsius incubator for 40 minutes. At the end of the incubation, place the tube back on the dissociator and run the Tumor 3 program for 37 seconds. At the end of the program, centrifuge the cells and re-suspend the pellet in 10 milliliters of serum free RPMI with gentle mixing.Next, slowly load the cells onto a 40 micron cell strainer in a new 50 milliliter tube, and allow the suspension to drip through the filter by gravity. Gently lift the tab of the cell strainer in between each new addition of cells, breaking the vacuum between the strainer and the tube to allow the suspended cells to pass freely through the mesh. When all of the cells have been filtered, spin down the single cell suspension and re-suspend the pellet in 10 milliliters of complete Neurobasal Medium.Then, determine the number of viable cells by trypan blue exclusion, and place the cells on ice. To generate the microtumors, spin down the dissociated patient derived glioblastoma xenoline cells and re-suspend the pellet at 50, 000 cells per two microliters of FBS. Next, dilute the cells in ice cold high density human biogel at a one to four cell to biogel ratio.Then, use an electronic multichannel pipette to dispense 10 microliters of cell mixture per pin onto a 96 pin steel plate with hydrophobic coating. It's important to prevent bubbles when mixing the cells, medium, and biogel, while still working quickly and efficiently to generate the microtumors before the human biogel begins to solidify on the pins. When all of the microtumors have been plated, transfer the 3D tumors into a tissue culture incubator for 20 minutes to gelate the tumor beads.At the end of the incubation, transfer the microtumors to a 10 centimeter culture dish containing complete Neurobasal Medium and return the tumors to the incubator. After one to two days, use a wide mouth dispensing pipette to transfer the microtumors into the wells of a 96 well culture plate, containing 50 microliters of Neurobasal Medium per well, and add 50 microliters of the appropriate dosing solution to each well. Maintain the treated microtumors in the tissue culture incubator for one to 14 days, feeding the tumors twice weekly with a fresh medium and drug solution.To evaluate the microtumor cell morphology, on days one, seven, and fourteen of culture, label the microtumors with one micromolar of freshly diluted Calcein-AM per well in the tissue culture incubator for 20 minutes. Then, image the tumors by flourescence microscopy at two, four, and 10x magnifications. To assess the microtumor growth, and days one, seven, and 14, treat the cultures with 20 microliters of freshly prepared MTT for two hours in the tissue culture incubator.At the end of the incubation, lyse the tumors with freshly prepared, 10%SDS lysis buffer at a one to one ratio to the culture well volumes, and seal the plate. Then incubate the microtumors in the lysis buffer overnight at 37 degrees Celsius, and read the absorbance at 570 nanometers on a multi-plate reader the next morning. Through live cell imaging Calcein-AM as just demonstrated, the cell growth and viability of the microtumors, as well as the effects of the drug therapies on these functions, can be evaluated.As observed, untreated microtumors and microtumors with a known resistance to the experimental drug treatment display a continuous growth pattern over a two week culture period. These TMZ sensitive tumor cells, however, demonstrate a reduced cell growth by the end of the culture in response to the highest concentration of TMZ treatment by MTT assay. Interestingly, kinase signaling in TMZ resistant microtumors reveals a number of peptides with significantly altered intensities after the 10 micromolar TMZ treatment.Indeed, in this representative experiment, analysis of these altered kinomic profiles using upstream kinase prediction software identified specific alterations in SYK, LCK, and CTK tyrosine kinase and a number of serine/threonine kinase intensities, suggesting potential novel therapeutic targets that may help overcome tumor treatment resistance. Once mastered, the tumor disaggregation can be completed in two hours if it is performed properly. The microtumor generation and drug dosing techniques can be completed in three hours total if they are performed properly.In the future, this model system could be used as a proband style avatar system, in which a patient's tumor is molecularly matched to an existing patient derived microtumor library to inform the clinician of the best choice of therapy.

Research

JoVE Journal

Medicine

Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester (CFSE)

Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester CFSE

और मानव Glioblastoma Carboxy Fluorescein Succinimidyl एस्टर (CFSE) में धीमी गति से विभाजित कोशिकाओं की पहचान अलगाव

Tumor heterogeneity represents a fundamental feature supporting tumor robustness and presents a central obstacle to the development of therapeutic ...

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

मानव दंत लुगदी स्टेम सेल के अलगाव विशेषता, और तुलनात्मक भेदभाव दो अलग अलग तरीकों का उपयोग करके स्थायी दांत से व्युत्पन्न

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human ...

Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma

Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma

रोगी व्युत्पन्न सेल संस्कृति और CD133 का अलगाव<sup&gt; +</sup&gt; मेलेनोमा से ख्यात कैंसर स्टेम सेल

Despite improved treatments options for melanoma available today, patients with advanced malignant melanoma still have a poor prognosis for ...

Generation of Prostate Cancer Patient Derived Xenograft Models from Circulating Tumor Cells

प्रोस्टेट कैंसर के रोगी की पीढ़ी परिसंचारी ट्यूमर कोशिकाओं से xenograft मॉडल व्युत्पन्न

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, ...

A Brain Tumor/Organotypic Slice Co-culture System for Studying Tumor Microenvironment and Targeted Drug Therapies

दवा के उपचारों ट्यूमर microenvironment अध्ययन और लक्षित के लिए एक मस्तिष्क ट्यूमर / Organotypic स्लाइस प्रणाली सह संस्कृति

Brain tumors are a major cause of cancer-related morbidity and mortality. Developing new therapeutics for these cancers is difficult, as many of these ...

Generation of a Humanized Mouse Liver Using Human Hepatic Stem Cells

एक humanized माउस जिगर की पीढ़ी मानव यकृत स्टेम कोशिकाओं का उपयोग

A novel animal model involving chimeric mice with humanized livers established via human hepatocyte transplantation has been developed. These mice, in ...

Drug Repurposing Hypothesis Generation Using the "RE:fine Drugs" System

ड्रग Repurposing परिकल्पना पीढ़ी &quot;उत्तर: ठीक ड्रग्स&quot; का उपयोग कर प्रणाली

The promise of drug repurposing is that existing drugs may be used for new disease indications in order to curb the high costs and time for approval. The ...

Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients

Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients

Forskolin प्रेरित आंतों में सूजन Organoids: एक<em&gt; इन विट्रो</em&gt; सिस्टिक फाइब्रोसिस रोगियों में दवा प्रतिक्रिया का आकलन करने के लिए परख

Recently-developed cystic fibrosis transmembrane conductance regulator (CFTR)-modulating drugs correct surface expression and/or function of the mutant ...

Assessment of the Anticoagulant and Anti-inflammatory Properties of Endothelial Cells Using 3D Cell Culture and Non-anticoagulated Whole Blood

3 डी सेल संस्कृति और गैर anticoagulated पूरे रक्त का उपयोग कर Endothelial कोशिकाओं के थक्कारोधी और विरोधी भड़काऊ गुणों का आकलन

In vivo, endothelial cells are crucial for the natural anticoagulation of circulating blood. Consequently, endothelial cell activation leads to blood ...

Muscle Imbalances: Testing and Training Functional Eccentric Hamstring Strength in Athletic Populations

मांसपेशी असंतुलन: परीक्षण और प्रशिक्षण कार्यात्मक विलक्षण दौडऩे पुष्ट आबादी में ताकत

Many hamstring injuries that occur during physical activity occur while the muscles are lengthening, during eccentric hamstring muscle actions. Opposite ...