Name: alginate microcapsule as a 3d platform for propagation and differentiation of human embryonic stem cells (hesc) to different lineages
Uploaded: 2012-03-09T12:00:00
Description: Watch this Scientific Journal Video about Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages at JoVE.com

यह काम NHMRC कार्यक्रम अनुदान +५६८९६९ # (PSS) से और संकाय मेडिसिन के न्यू साउथ वेल्स विश्वविद्यालय, स्टेम सेल पहल (KSS) के द्वारा समर्थित है.

नीचे सभी प्रक्रियाओं की एक द्वितीय श्रेणी जैवसुरक्षा मंत्रिमंडल के अंदर बाहर किया जाता है सड़न रोकनेवाला तकनीक का उपयोग कर. अभिकर्मकों और उपकरणों का इस्तेमाल किया नीचे दी गई तालिका में सूचीबद्ध हैं. ...

<ol>
	<li>Chayosumrit, M., Tuch, B., Sidhu, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alginate+microcapsule+for+propagation+and+directed+differentiation+of+hESCs+to+definitive+endoderm.">Alginate microcapsule for propagation and directed differentiation of hESCs to definitive endoderm.</a> Biomaterials. 31, 505-514 (2010).</li><li>Dean, S. K., Yulyana, Y., Williams, G., Sidhu, K. S., Tuch, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differentiation+of+encapsulated+embryonic+stem+cells+after+transplantation.">Differentiation of encapsulated embryonic stem cells after transplantation.</a> Transplantation. 82, 1175-1184 (2006).</li><li>Siti-Ismail, N., Bishop, A. E., Polak, J. M., Mantalaris, 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+benefit+of+human+embryonic+stem+cell+encapsulation+for+prolonged+feeder-free+maintenance.">The benefit of human embryonic stem cell encapsulation for prolonged feeder-free maintenance.</a> Biomaterials. 29, 3946-3952 (2008).</li><li>Dawson, E., Mapili, G., Erickson, K., Taqvi, S., Roy, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomaterials+for+stem+cell+differentiation.">Biomaterials for stem cell differentiation.</a> Adv. Drug Deliv. Rev. 60, 215-228 (2008).</li><li>Orive, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+encapsulation:+promise+and+progress.">Cell encapsulation: promise and progress.</a> Nat. Med. 9, 104-107 (2003).</li><li>Watanabe, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+ROCK+inhibitor+permits+survival+of+dissociated+human+embryonic+stem+cells.">A ROCK inhibitor permits survival of dissociated human embryonic stem cells.</a> Nat. Biotechnol. 25, 681-686 (2007).</li><li>Dang, S. M., Gerecht-Nir, S., Chen, J., Itskovitz-Eldor, J., Zandstra, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled,+scalable+embryonic+stem+cell+differentiation+culture.">Controlled, scalable embryonic stem cell differentiation culture.</a> Stem Cells. 22, 275-282 (2004).</li><li>Drukker, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+expression+of+MHC+proteins+in+human+embryonic+stem+cells.">Characterization of the expression of MHC proteins in human embryonic stem cells.</a> Proc. Natl. Acad. Sci. U S A. 99, 9864-9869 (2002).</li><li>Calafiore, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standard+Technical+Procedures+for+Microencapsulation+of+Human+Islets+for+Graft+into+Nonimmunosuppressed+Patients+With+Type+1+Diabetes+Mellitus.">Standard Technical Procedures for Microencapsulation of Human Islets for Graft into Nonimmunosuppressed Patients With Type 1 Diabetes Mellitus.</a> Transplantation Proceedings. 38, 1156-1157 (2006).</li><li>Cho, M. S. <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+and+large-scale+generation+of+functional+dopamine+neurons+from+human+embryonic+stem+cells.">Highly efficient and large-scale generation of functional dopamine neurons from human embryonic stem cells.</a> Proc. Natl. Acad. Sci. U.S.A. 105, 3392-3397 (2008).</li><li>Vazin, T., Chen, J., Lee, C. T., Amable, R., Freed, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+stromal-derived+inducing+activity+in+the+generation+of+dopaminergic+neurons+from+human+embryonic+stem+cells.">Assessment of stromal-derived inducing activity in the generation of dopaminergic neurons from human embryonic stem cells.</a> Stem Cells. 26, 1517-1525 (2008).</li></ol>

कई माउस भ्रूणीय स्टेम कोशिकाओं और hESC का उपयोग अध्ययन biomaterials और ऊतक इंजीनियरिंग 2,3 में 3 डी संस्कृति प्रणाली के लाभ का प्रदर्शन किया है. हम एक उपयुक्त 3 डी मंच के रूप में बेरियम alginate की तुलना में hESC प्रचार और...

<table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td><td> नोट्स </td></tr><tr><td> Alginate (MVG उत्तर प्रदेश ProNova) </td><td> NovoMatrix </td><td> 4200101 </td><td> उच्च glucuronic एसिड ≥ 60% सामग्री, दलदलापन&gt; 200 MPa है, और अन्तर्जीवविष &lt;100 यूरोपीय संघ / छ </td></tr><tr><td> जेलाटीन </td><td> सिग्मा Aldrich </td><td> G1890-100g </td><td></td></tr><tr><td> 0.9% NaCl </td><td> बैक्सटर स्वास्थ्य सेवा </td><td> AHF7123 </td><td></td></tr><tr><td> प्रकार J1 मनका जनरेटर </td><td> Nisco इंजीनियरिंग इंक </td><td> SPA-0447 </td><td></td></tr><tr><td> बहु - Phaser सिरिंज पंप </td><td> नए युग पम्प सिस्टम्स इंक </td><td> पूर्वोत्तर के एक हज़ार मॉडल </td><td></td></tr><tr><td> Ezi फ्लो चिकित्सा Flowmeter </td><td> डींगमार सिस्टम</td><td> G0149 </td><td></td></tr><tr><td> Y - 27,632 </td><td> मर्क </td><td> 688.000 </td><td></td></tr><tr><td> सीरम मानव Albumin </td><td> सिग्मा Aldrich </td><td> A4327-1G </td><td></td></tr><tr><td> Accutase </td><td> Millipore </td><td> SCR005 </td><td></td></tr><tr><td> 14G एक्स 2 &quot;चतुर्थ कैथेटर </td><td> Terumo </td><td> SR-OX1451C </td><td></td></tr><tr><td> नॉकआउट - DMEM </td><td> Invitrogen </td><td> 10829-018 </td><td> एसआर मध्यम (बेसल) के लिए </td></tr><tr><td> GlutaMAX मैं </td><td> Invitrogen </td><td> 35050-061 </td><td> एसआर मध्यम (2 मिमी) के लिए </td></tr><tr><td> नॉकआउट सीरम रिप्लेसमेंट </td><td> Invitrogen </td><td> 10828-028 </td><td> एसआर मध्यम (20%) के लिए </td></tr><tr><td> पेनिसिलिन - स्ट्रेप्टोमाइसिन </td><td> Invitrogen </td><td> 15070063 </td><td> एसआर मध्यम (2.5 मिलीग्राम / यू) के लिए </td&gt; </tr><tr><td> इंसुलिन transferrin - सेलेनियम (आईटीएस) </td><td> Invitrogen </td><td> 41400045 </td><td> एसआर मध्यम (1x) </td></tr><tr><td> β-Mercaptoethanol </td><td> Invitrogen </td><td> 21985-023 </td><td> एसआर मध्यम (0.1 मिमी) के लिए </td></tr><tr><td> सदस्य NEAA समाधान </td><td> Invitrogen </td><td> 11140-050 </td><td> एसआर मध्यम (5 मिमी) के लिए </td></tr><tr><td> ग्लासगो न्यूनतम आवश्यक मध्यम </td><td> Invitrogen </td><td> 11710035 </td><td> डीए तंत्रिका भेदभाव मध्यम (बेसल) के लिए </td></tr><tr><td> नॉकआउट सीरम रिप्लेसमेंट </td><td> Invitrogen </td><td> 10828-028 </td><td> डीए तंत्रिका भेदभाव मध्यम (10%) के लिए </td></tr><tr><td> सोडियम पाइरूवेट </td><td> Invitrogen </td><td> 11360070 </td><td> डीए तंत्रिका भेदभाव मध्यम (1 मिमी) के लिए </td></tr><tr><td> सदस्य NEAA समाधान </td><td> Invitrogen </td><td> 11140-050 </td><td> डीए पता के लिएeural भेदभाव मध्यम (0.1 मिमी) </td></tr><tr><td> β-Mercaptoethanol </td><td> Invitrogen </td><td> 21985-023 </td><td> डीए तंत्रिका भेदभाव मध्यम (0.1 मिमी) के लिए </td></tr><tr><td> ध्वनि का हाथी (श) </td><td> अनुसंधान एवं विकास सिस्टम </td><td> 1314-SH-025/CF </td><td> डीए तंत्रिका भेदभाव (100 एनजी / मिली) के लिए </td></tr><tr><td> Fibroblast वृद्धि कारक 8a (FGF8a) </td><td> अनुसंधान एवं विकास सिस्टम </td><td> ४७४५-F8-050 </td><td> डीए तंत्रिका भेदभाव (100 एनजी / मिली) के लिए </td></tr></tbody></table>

alginate microcapsule as a 3d platform for propagation and differentiation of human embryonic stem cells (hesc) to different lineages

Human embryonic stem cells (hESC) are emerging as an attractive alternative source for cell replacement therapy since they can be expanded in culture indefinitely and differentiated to any cell types in the body. Various types of biomaterials have also been used in stem cell cultures to provide a microenvironment mimicking the stem cell niche1-3. The latter is important for promoting cell-to-cell interaction, cell proliferation, and differentiation into specific lineages as well as tissue organization by providing a three-dimensional (3D) environment4 such as encapsulation. The principle of cell encapsulation involves entrapment of living cells within the confines of semi-permeable membranes in 3D cultures2. These membranes allow for the exchange of nutrients, oxygen and stimuli across the membranes, whereas antibodies and immune cells from the host that are larger than the capsule pore size are excluded5. Here, we present an approach to culture and differentiate hESC DA neurons in a 3D microenvironment using alginate microcapsules. We have modified the culture conditions2 to enhance the viability of encapsulated hESC. We have previously shown that the addition of p160-Rho-associated coiled-coil kinase (ROCK) inhibitor, Y-27632 and human fetal fibroblast-conditioned serum replacement medium (hFF-CM) to the 3D platform significantly enhanced the viability of encapsulated hESC in which the cells expressed definitive endoderm marker genes1. We have now used this 3D platform for the propagation of hESC and efficient differentiation to DA neurons. Protein and gene expression analyses after the final stage of DA neuronal differentiation showed an increased expression of tyrosine hydroxylase (TH), a marker for DA neurons, &gt;100 folds after 2 weeks. We hypothesized that our 3D platform using alginate microcapsules may be useful to study the proliferation and directed differentiation of hESC to various lineages. This 3D system also allows the separation of feeder cells from hESC during the process of differentiation and also has potential for immune-isolation during transplantation in the future.

Watch this Scientific Journal Video about Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages at JoVE.com

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages

हम प्रचार और अन्तर्जनस्तर और डोपामिनर्जिक न्यूरॉन्स (डीए) को भ्रूण स्टेम कोशिकाओं की भिन्नता के लिए एक प्रभावी 3D मंच के रूप में एक microencapsulation तकनीक अनुकूलित किया. यह भी प्रतिरक्षा कोशिकाओं के प्रत्यारोपण के दौरान मेजबान से अलगाव के लिए एक अवसर प्रदान करता है. इस मंच के अन्य प्रकार की कोशिकाओं के लिए अनुकूलित किया जा सकता है.

और मानव भ्रूण स्टेम कोशिकाओं (hESC) के प्रसार और भेदभाव के लिए एक 3 डी मंच के रूप में विभिन्न प्रजातियों के लिए alginate microcapsule

Human embryonic stem cells (hESC) are emerging as an attractive alternative source for cell replacement therapy since they can be expanded in culture ...

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Bioengineering

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells (hESC) to Different Lineages

Rotating Cell Culture Systems for Human Cell Culture: Human Trophoblast Cells as a Model

मानव सेल संस्कृति घूर्णन सेल संस्कृति सिस्टम: एक मॉडल के रूप में मानव ट्रोफोब्लास्ट कक्ष

The field of human trophoblast research aids in  understanding the complex environment established during placentation. Due to the  nature of these ...

Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells (hESCs) and Induced Pluripotent Stem Cells (iPSCs)

Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells hESCs and Induced Pluripotent Stem Cells iPSCs

विकास, विस्तार, और<em&gt; विवो में</em&gt; मानव भ्रूण स्टेम कोशिकाओं (hESCs) से मानव एन.के. कोशिकाओं की निगरानी और और प्रेरित pluripotent स्टेम सेल (iPSCs)

We present a method for deriving natural killer (NK) cells from undifferentiated hESCs and iPSCs using a feeder-free approach. This method gives rise to ...

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एकल कक्षों और 3 डी कारावास में एकल भ्रूण आदेश: उच्च सामग्री स्क्रीनिंग के लिए एक नई डिवाइस

Biological cells are usually observed on flat (2D) surfaces. This condition is not physiological, and phenotypes and shapes are highly variable. Screening ...

Layered Alginate Constructs: A Platform for Co-culture of Heterogeneous Cell Populations

बहुस्तरीय Alginate constructs: विषम सेल आबादी के सह संस्कृति के लिए एक मंच

Many load bearing tissues possess structurally and functionally distinct regions, typically accompanied by different cell phenotypes with differential ...

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Micropatterning और 3 डी microvessels के विधानसभा

In vitro platforms to study endothelial cells and vascular biology are largely limited to 2D endothelial cell culture, flow chambers with polymer or glass ...

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मानव प्रेरित Pluripotent स्टेम सेल-व्युत्पन्न कार्डियोमायोसाइट्स में एकल सेल ऑप्टिकल एक्शन संभावित माप

Conventional intracellular microelectrode techniques to quantify cardiomyocyte electrophysiology are extremely complex, labor intensive, and typically ...

A Culture Method to Maintain Quiescent Human Hematopoietic Stem Cells

शांत मानव हेमेटोपोइटिक स्टेम सेल को बनाए रखने के लिए एक संस्कृति विधि

Human hematopoietic stem cells (HSCs), like other mammalian HSCs, maintain lifelong hematopoiesis in the bone marrow. HSCs remain quiescent in vivo, ...

Real-Time Intravital Multiphoton Microscopy to Visualize Focused Ultrasound and Microbubble Treatments to Increase Blood-Brain Barrier Permeability

वास्तविक समय Intravital Multiphoton माइक्रोस्कोपी केंद्रित अल्ट्रासाउंड और Microbubble उपचार की कल्पना करने के लिए रक्त मस्तिष्क बाधा पारगम्यता बढ़ाने के लिए

The blood-brain barrier (BBB) is a key challenge for the successful delivery of drugs to the brain. Ultrasound exposure in the presence of microbubbles ...

In vitro Induction of Human Dental Pulp Stem Cells Toward Pancreatic Lineages

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इन विट्रो अग्नाशय के वंश की ओर मानव दंत लुगदी स्टेम सेल का प्रेरण

As of 2000, the success of pancreatic islet transplantation using the Edmonton protocol to treat type I diabetes mellitus still faced some obstacles. ...

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माइक्रोफ्लुइडिक प्लेटफॉर्म के भीतर 3डी संगठित मानव हृदय ऊतक का विकास

The leading cause of death worldwide persists as cardiovascular disease (CVD). However, modeling the physiological and biological complexity of the heart ...