Name: directed differentiation of induced pluripotent stem cells towards t lymphocytes
Uploaded: 2012-05-14T13:00:00
Description: Watch this Scientific Journal Video about Directed Differentiation of Induced Pluripotent Stem Cells towards T Lymphocytes at JoVE.com

हम प्रदान करने के लिए डॉ. Shinya Yamanaka (क्योटो विश्वविद्यालय) धन्यवाद आईपीएस MEF एनजी 20D-17 सेल लाइन का समर्थन करने के लिए OT1-2A डॉ. Dario Vignali (सेंट जूदास बच्चों अनुसंधान अस्पताल) • pMig द्वितीय का निर्माण, डा. जुआन कार्लोस Zuniga सेल OP9 DL1 लाइन का समर्थन करने के लिए (इम्यूनोलॉजी विभाग, टोरंटो विश्वविद्यालय के) - Pflucker, और इस अध्ययन के डिजाइन की मदद करने के लिए डॉ. केंट ई Vrana (औषधि विभाग, पेन स्टेट यूनिवर्सिटी कॉलेज ऑफ मेडिसिन के). इस परियोजना, राष्ट्रीय कैंसर संस्थान, Barsumian ट्रस्ट और मेलेनोमा रिसर्च फाउंडेशन (जे सांग) से अनुदान K18CA151798 संख्या के साथ अनुदान के तहत वित्त पोषित है.

1. सेल संस्कृति <ol><li> / 7 SNL76 विकिरणित (/ 7 irSNL76) संस्कृति के लिए फीडर कोशिकाओं की तैयारी. SNL76 / 7 कोशिकाओं आम तौर पर 10% भ्रूण गोजातीय सीरम (FBS) Dulbecco संशोधित ईगल मध्यम (DMEM) मीडिया में रखा जाता है. <ol><li> तरल नाइट्रोजन से SNL76 / 7 क?...

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<html> अधिनियम आधारित चिकित्सा के लिए, पुनः जलसेक vivo में अत्यधिक प्रतिक्रियाशील एजी विशेष टी कोशिकाओं की बड़ी संख्या की इन विट्रो पीढ़ी में एक इष्टतम दृष्टिकोण है. हालांकि हमारे विधि में इन विट...

<table border="1"><tbody><tr><td> अभिकर्मक के नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td></tr><tr><td> C57BL/6J चूहों </td><td> जैक्सन प्रयोगशाला </td><td> 000664 </td></tr><tr><td> B6.129S7 - Rag1 tm1Mom जम्मू / </td><td> जैक्सन प्रयोगशाला </td><td> 002216 </td></tr><tr><td> विरोधी CD3 प्रतिरक्षी (2C11) </td><td> BD PharMingen </td><td> 553058 </td></tr><tr><td> विरोधी CD28 एंटीबॉडी (37.51) </td><td> BD PharMingen </td><td> 553295 </td></tr><tr><td> विरोधी CD3 प्रतिरक्षी (17A2) </td><td> BioLegend </td><td> 100202 </td></tr><tr><td> विरोधी CD4 (GK1.5) एंटीबॉडी </td><td> BioLegend </td><td> 100417 </td></tr><tr><td> (53-6.7) विरोधी CD8 एंटीबॉडी </td><td> BioLegend </td><td> 100714 </td></tr><tr><td> विरोधी CD25 एंटीबॉडी (3C7) </td><td> BioLegend </td><td> 101912 </td></tr><tr><td> एकती CD44 एंटीबॉडी (1M7) </td><td> BioLegend </td><td> 103012 </td></tr><tr><td> विरोधी CD117 एंटीबॉडी (2B8) </td><td> BioLegend </td><td> 105812 </td></tr><tr><td> विरोधी TCR β (H57597) एंटीबॉडी </td><td> BioLegend </td><td> 109220 </td></tr><tr><td> विरोधी (JES6 5H4) IL-2 एंटीबॉडी </td><td> BioLegend </td><td> 503810 </td></tr><tr><td> विरोधी IFN-γ एंटीबॉडी (XMG1.2) </td><td> BioLegend </td><td> 505822 </td></tr><tr><td> DMEM </td><td> Invitrogen </td><td> ABCD1234 </td></tr><tr><td> α - सदस्य </td><td> Invitrogen </td><td> A10490-01 </td></tr><tr><td> FBS </td><td> HyClone </td><td> SH3007.01 </td></tr><tr><td> एक Brefeldin </td><td> सिग्मा </td><td> B7651 </td></tr><tr><td> Polybrene </td><td> सिग्मा </td><td> 107689 </td></tr><tr><td> GeneJammer </td><td> एकीकृत विज्ञान </td><td> 204130 </td></tr><tr><td> आरएनए किट &lt;/ P&gt; <td> क्विएज़न </td><td> 74104 </td></tr><tr><td> डीएनए किट </td><td> क्विएज़न </td><td> 69504 </td></tr><tr><td> CD8 अलगाव किट </td><td> Miltenyi बायोटेक </td><td> 130-095-236 </td></tr><tr><td> ACK lysis बफर </td><td> LONZA </td><td> 10-548E </td></tr><tr><td> mFlt-3L </td><td> PeproTech </td><td> 250-31L </td></tr><tr><td> 7 लाख </td><td> PeproTech </td><td> 217-17 </td></tr><tr><td> जेलाटीन </td><td> सिग्मा </td><td> G9391 </td></tr><tr><td> FITC विरोधी OVA एंटीबॉडी </td><td> रॉकलैंड Immunochemicals </td><td> 200-4233 </td></tr><tr><td> Permeabilization बफर </td><td> Biolegend </td><td> 421002 </td></tr><tr><td> BSA </td><td> सिग्मा </td><td> A7906 </td></tr><tr><td> नि: संक्रामक </td><td> सिग्मा </td><td> F8775 </td></tr><tr><td> 0.4 फिल्टर सुक्ष्ममापी </td><td> MILLIPORE </td><td></td></tr><tr><td&gt; Moflo सेल सॉर्टर </td><td> Dake Cytomation </td><td></td></tr><tr><td> Calibur प्रवाह cytometer </td><td> BD </td><td></td></tr><tr><td> LSR द्वितीय प्रवाह cytometer </td><td> BD </td><td></td></tr><tr><td> माउस restrainer </td><td> Braintree वैज्ञानिक </td><td></td></tr></tbody></table>

directed differentiation of induced pluripotent stem cells towards t lymphocytes

Adoptive cell transfer (ACT) of antigen-specific CD8+ cytotoxic T lymphocytes (CTLs) is a promising treatment for a variety of malignancies 1. CTLs can recognize malignant cells by interacting tumor antigens with the T cell receptors (TCR), and release cytotoxins as well as cytokines to kill malignant cells. It is known that less-differentiated and central-memory-like (termed highly reactive) CTLs are the optimal population for ACT-based immunotherapy, because these CTLs have a high proliferative potential, are less prone to apoptosis than more differentiated cells and have a higher ability to respond to homeostatic cytokines 2-7. However, due to difficulties in obtaining a high number of such CTLs from patients, there is an urgent need to find a new approach to generate highly reactive Ag-specific CTLs for successful ACT-based therapies.
TCR transduction of the self-renewable stem cells for immune reconstitution has a therapeutic potential for the treatment of diseases 8-10. However, the approach to obtain embryonic stem cells (ESCs) from patients is not feasible. Although the use of hematopoietic stem cells (HSCs) for therapeutic purposes has been widely applied in clinic 11-13, HSCs have reduced differentiation and proliferative capacities, and HSCs are difficult to expand in in vitro cell culture 14-16. Recent iPS cell technology and the development of an in vitro system for gene delivery are capable of generating iPS cells from patients without any surgical approach. In addition, like ESCs, iPS cells possess indefinite proliferative capacity in vitro, and have been shown to differentiate into hematopoietic cells. Thus, iPS cells have greater potential to be used in ACT-based immunotherapy compared to ESCs or HSCs.
Here, we present methods for the generation of T lymphocytes from iPS cells in vitro, and in vivo programming of antigen-specific CTLs from iPS cells for promoting cancer immune surveillance. Stimulation in vitro with a Notch ligand drives T cell differentiation from iPS cells, and TCR gene transduction results in iPS cells differentiating into antigen-specific T cells in vivo, which prevents tumor growth. Thus, we demonstrate antigen-specific T cell differentiation from iPS cells. Our studies provide a potentially more efficient approach for generating antigen-specific CTLs for ACT-based therapies and facilitate the development of therapeutic strategies for diseases.

Watch this Scientific Journal Video about Directed Differentiation of Induced Pluripotent Stem Cells towards T Lymphocytes at JoVE.com

Directed Differentiation of Induced Pluripotent Stem Cells towards T Lymphocytes

प्रेरित pluripotent स्टेम सेल (आईपीएस) से टी lymphocytes की पीढ़ी टी immunotherapy सेल आधारित के लिए भ्रूण स्टेम कोशिकाओं का उपयोग करने का एक वैकल्पिक दृष्टिकोण देता है. विधि से पता चलता है कि या तो द्वारा का उपयोग<em&gt; इन विट्रो में</em&gt; या<em&gt; Vivo में</em&gt; प्रेरण सिस्टम, आईपीएस कोशिकाओं दोनों पारंपरिक और प्रतिजन विशेष टी लिम्फोसाइटों में अंतर करने में सक्षम हैं.

टी lymphocytes की ओर प्रेरित pluripotent स्टेम सेल की निर्देशित भेदभाव

Adoptive cell transfer (ACT) of antigen-specific CD8+  cytotoxic T lymphocytes (CTLs) is a promising treatment for a variety of  malignancies 1. CTLs can  ...

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साइटोटोक्सिक टी lymphocytes द्वारा बीटा सेल मौत मध्यस्थता का आकलन करने के तरीके

Type 1 diabetes (T1D) is a T cell mediated autoimmune disease.  During the pathogenesis, patients become progressively more insulinopenic as 
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Virus infections after stem cell transplantation are among the most common causes of death, especially after cord blood (CB) transplantation (CBT) where ...

Generation of Induced Regulatory T Cells from Primary Human Na&iuml;ve and Memory T Cells

Generation of Induced Regulatory T Cells from Primary Human NaÃƒÆ’Ã‚Â¯ve and Memory T Cells

प्राथमिक मानव भोले और स्मृति टी कोशिकाओं से प्रेरित विनियामक टी कोशिकाओं की पीढ़ी

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अनुभवहीन सीडी 4 की adenoviral पारगमन Treg भेदभाव का अध्ययन करने के लिए कोशिकाओं टी

Regulatory T cells (Tregs) are essential to provide immune tolerance to self as well as to certain foreign antigens. Tregs can be generated from naive CD4 ...

Isolation and Th17 Differentiation of Na&iuml;ve CD4 T Lymphocytes

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Th17 cells are a distinct subset of T cells that have been found to produce interleukin 17 (IL-17), and differ in function from the other T cell subsets ...

Derivation of T Cells In Vitro from Mouse Embryonic Stem Cells

Derivation of T Cells In Vitro from Mouse Embryonic Stem Cells

टी कोशिकाओं की व्युत्पत्ति<em&gt; इन विट्रो</em&gt; माउस भ्रूणीय स्टेम कोशिकाओं से

The OP9/OP9-DL1 co-culture system has become a well-established method for deriving differentiated blood cell types from embryonic and hematopoietic ...

In Vitro Differentiation of Human CD4+FOXP3+ Induced Regulatory T Cells (iTregs) from Na&#239;ve CD4+ T Cells Using a TGF-&#946;-containing Protocol

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भोले सीडी 4 + टी कोशिकाओं से मानव सीडी 4 + Foxp3 + प्रेरित विनियामक टी कोशिकाओं (iTregs) एक TGF-β युक्त प्रोटोकॉल का उपयोग की इन विट्रो भेदभाव में

Regulatory T cells (Tregs) are an integral part of peripheral tolerance, suppressing immune reactions against self-structures and thus preventing ...

Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs

इम्यूनोमॉड्यूलेटरी औषध पहचान के लिए टी lymphocytes में माइटोजन प्रेरित Blastogenesis की रैपिड मात्रा

Lymphocyte proliferation in response to antigenic or mitogenic stimulation is a readily quantifiable phenomenon useful for testing immunomodulatory (i.e., ...

An In Vivo Mouse Model to Measure Na&#239;ve CD4 T Cell Activation, Proliferation and Th1 Differentiation Induced by Bone Marrow-derived Dendritic Cells

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एक Vivo माउस मॉडल में भोली CD4 टी सेल सक्रियण, प्रसार और Th1 अस्थि मज्जा द्वारा प्रेरित भेदभाव-व्युत्पंन वृक्ष कोशिकाओं को मापने के लिए

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