Name: preparation of 3d fibrin scaffolds for stem cell culture applications
Uploaded: 2012-03-02T12:00:00
Description: Watch this Scientific Journal Video about Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications at JoVE.com

लेखकों के लिए प्रेरित pluripotent स्टेम सेल के व्यवहार को नियंत्रित करने के लिए ऊतक इंजीनियर scaffolds के NSERC डिस्कवरी अनुदान 402,462 स्वीकार करते हैं करना चाहते हैं.

प्रोटोकॉल शुरू करने से पहले नोट: फाइब्रिनोजेन एक रक्त प्रोटीन व्युत्पन्न है और इस प्रकार उचित सुरक्षा प्रशिक्षण से निपटने से पहले पूरा किया जाना चाहिए. इस प्रोटोकॉल के 2 दिन के समय तो वांछित स्ट?...

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इस प्रोटोकॉल के ऊपर विस्तृत pluripotent स्टेम सेल संस्कृति के लिए 3 डी आतंच scaffolds के सृजन, माउस भ्रूण और प्रेरित pluripotent स्टेम सेल के लिए विशेष रूप से करने के लिए एक तरीका प्रदान करता है. इस 3 डी biomaterial आधारित संस्कृति प्?...

उपकरण की जरूरत है: एनालाइटिकल बैलेंस पीएच मीटर टिशू कल्चर इनक्यूबेटर (37 डिग्री सेल्सियस, 5% सीओ 2) थाली हलचल स्पेक्ट्रोफोटोमीटर बाँझ ऊतक संस्कृति हुड Tris खारा बफर (7.4 पीएच) (प्लस पर्याप्त फाइब्रिनोजेन भंग करने के लिए की आवश्यकता है 4 एल) 50 मिमी कैल्शियम क्लोराइड समाधान बाँझ चोटीदार ट्यूबों (15 या 50 एमएल) 35 मिमी पेट्री डिश डायलिसिस टयूबिंग (+७,००० मेगावाट cutoff) डायलिसिस क्लिप 5.0 माइक्रोन सिरिंज फिल्टर व्यक्तिगत लिपटे बाँझ 0.22 माइक्रोन सिरिंज फिल्टर सिरिंज 24 अच्छी तरह से ऊतक संस्कृति प्लेट <table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td><td> टिप्पणियाँ </td></tr><tr><td> फाइब्रिनोजेन (मानव) </td><td> Calbiocहेम </td><td> 341,578 </td><td></td></tr><tr><td> थ्रोम्बिन (मानव) </td><td> सिग्मा </td><td> T7009 </td><td></td></tr></tbody></table>

preparation of 3d fibrin scaffolds for stem cell culture applications

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells inside of 3D biomaterial scaffolds provides a way to accurately mimic these microenvironments, providing an advantage over traditional 2D culture methods using polystyrene as well as a method for engineering replacement tissues 2. While 2D tissue culture polystrene has been used for the majority of cell culture experiments, 3D biomaterial scaffolds can more closely replicate the microenvironments found in vivo by enabling more accurate establishment of cell polarity in the environment and possessing biochemical and mechanical properties similar to soft tissue.3 A variety of naturally derived and synthetic biomaterial scaffolds have been investigated as 3D environments for supporting stem cell growth. While synthetic scaffolds can be synthesized to have a greater range of mechanical and chemical properties and often have greater reproducibility, natural biomaterials are often composed of proteins and polysaccharides found in the extracelluar matrix and as a result contain binding sites for cell adhesion and readily support cell culture. Fibrin scaffolds, produced by polymerizing the protein fibrinogen obtained from plasma, have been widely investigated for a variety of tissue engineering applications both in vitro and in vivo 4. Such scaffolds can be modified using a variety of methods to incorporate controlled release systems for delivering therapeutic factors 5. Previous work has shown that such scaffolds can be used to successfully culture embryonic stem cells and this scaffold-based culture system can be used to screen the effects of various growth factors on the differentiation of the stem cells seeded inside 6,7.
 This protocol details the process of polymerizing fibrin scaffolds from fibrinogen solutions using the enzymatic activity of thrombin. The process takes 2 days to complete, including an overnight dialysis step for the fibrinogen solution to remove citrates that inhibit polymerization. These detailed methods rely on fibrinogen concentrations determined to be optimal for embryonic and induced pluripotent stem cell culture. Other groups have further investigated fibrin scaffolds for a wide range of cell types and applications - demonstrating the versatility of this approach 8-12.

Watch this Scientific Journal Video about Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications at JoVE.com

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

यह काम 3 डी आतंच की तैयारी विवरण के संवर्धन और plutipotent स्टेम कोशिकाओं फर्क करने के लिए scaffolds. इस तरह के scaffolds के स्टेम सेल के व्यवहार पर विभिन्न जैविक यौगिकों के प्रभाव स्क्रीन के रूप में अच्छी तरह के रूप में दवा वितरण प्रणाली को शामिल करने के लिए संशोधित करने के लिए इस्तेमाल किया जा सकता है.

3 डी आतंच की तैयारी स्टेम सेल संस्कृति अनुप्रयोगों के लिए scaffolds

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells ...

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Bioengineering

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

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

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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Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting ...

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Three dimensional cell cultures are attractive models for biological research. They combine the flexibility and cost-effectiveness of cell culture with ...

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Human articular cartilage is highly susceptible to damage and has limited self-repair and regeneration potential. Cell-based strategies to engineer ...

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उपकला सेल repopulation और गुर्दे ऊतकों के विकास के लिए कृंतक बाह्य मैट्रिक्स scaffolds के तैयारी

This protocol details the generation of acellular, yet biofunctional, renal extracellular matrix (ECM) scaffolds that are useful as small-scale model ...

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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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सेल संस्कृति अनुप्रयोगों के लिए प्रकाश की मध्यस्थता गठन और Hydrogels के patterning

Click chemistries have been investigated for use in numerous biomaterials applications, including drug delivery, tissue engineering, and cell culture. In ...

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3 डी स्थानिक सेल संस्कृतियों के लिए सेल Scaffolds के रूप में Biocompatible लिक्विड क्रिस्टल Elastomer Foams का संश्लेषण

Here, we present a step-by-step preparation of a 3D, biodegradable, foam-like cell scaffold. These scaffolds were prepared by cross-linking star block ...

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एक 3 डी गतिशील संस्कृति प्रणाली के साथ एक Multilayered Mesenchymal स्टेम सेल शीट का निर्माण

Stem cell therapy shows a promising future in regenerating injured organ and tissues, and the cell sheet technique has been developed to improve the low ...

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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, ...