Summary

एक माउस के मॉडल Utero में ट्रांसप्लांटेशन

Published: January 27, 2011
doi:

Summary

माउस के मॉडल<em> Utero में</em> प्रत्यारोपण एक बहुमुखी उपकरण है कि भ्रूण में स्टेम सेल और जीन थेरेपी प्रत्यारोपण के संभावित नैदानिक ​​अनुप्रयोगों का अध्ययन करने के लिए इस्तेमाल किया जा सकता है. इस प्रोटोकॉल में, हम इस तकनीक का प्रदर्शन करने के लिए एक सामान्य दृष्टिकोण प्रस्तुत

Abstract

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus. For example, in utero transplantation (IUT) of hematopoietic stem cells has been used to successfully treat patients with severe combined immunodeficiency.1,2 In several other conditions, however, IUT has been attempted without success.3 Given these mixed results, the availability of an efficient non-human model to study the biological sequelae of stem cell transplantation and gene therapy is critical to advance this field. We and others have used the mouse model of IUT to study factors affecting successful engraftment of in utero transplanted hematopoietic stem cells in both wild-type mice4-7 and those with genetic diseases.8,9 The fetal environment also offers considerable advantages for the success of in utero gene therapy. For example, the delivery of adenoviral10, adeno-associated viral10, retroviral11, and lentiviral vectors12,13 into the fetus has resulted in the transduction of multiple organs distant from the site of injection with long-term gene expression. in utero gene therapy may therefore be considered as a possible treatment strategy for single gene disorders such as muscular dystrophy or cystic fibrosis. Another potential advantage of IUT is the ability to induce immune tolerance to a specific antigen. As seen in mice with hemophilia, the introduction of Factor IX early in development results in tolerance to this protein.14

In addition to its use in investigating potential human therapies, the mouse model of IUT can be a powerful tool to study basic questions in developmental and stem cell biology. For example, one can deliver various small molecules to induce or inhibit specific gene expression at defined gestational stages and manipulate developmental pathways. The impact of these alterations can be assessed at various timepoints after the initial transplantation. Furthermore, one can transplant pluripotent or lineage specific progenitor cells into the fetal environment to study stem cell differentiation in a non-irradiated and unperturbed host environment.

The mouse model of IUT has already provided numerous insights within the fields of immunology, and developmental and stem cell biology. In this video-based protocol, we describe a step-by-step approach to performing IUT in mouse fetuses and outline the critical steps and potential pitfalls of this technique.

Protocol

1. इंजेक्शन pipettes तैयार जैसे कि कांच विंदुक की जुदाई 15 सेकंड (निर्माता अंशांकन के बारे में निर्देश देखने के) के भीतर होता है विंदुक खींचने जांचना. विंदुक एक घटना जहां यह अलग होगा. ऐसी है कि घटना की शु?…

Discussion

से अधिक 50 साल पहले, billingham, ब्रेंट, और Medawar चूहों में utero प्रत्यारोपण में इस्तेमाल के लिए विदेशी प्रोटीन के लिए प्रतिरक्षा सहिष्णुता प्रेरित उस समय के बाद से. 16, इस तकनीक के कई रूपों इम्यूनोलॉजी और स?…

Disclosures

The authors have nothing to disclose.

Acknowledgements

हम हमारे वित्तपोषण स्रोतों को स्वीकार करना चाहते हैं: कैलिफोर्निया इंस्टीट्यूट ऑफ पुनर्योजी चिकित्सा क्लीनिकल फैलो प्रशिक्षण अनुदान के लिए (एक), राष्ट्रीय विज्ञान फाउंडेशन (मेगावाट), Irene Perstein पुरस्कार (TCM), सर्जन के अमेरिकी कॉलेज (TCM), अमेरिकी बाल चिकित्सा सर्जिकल एसोसिएशन ( टीसीएम), और ऑफ डाइम्स (टीसीएम) के मार्च.

Materials

Material Name Type Company Catalogue Number Comment
Pipettes   Kimble 71900-100  
Pipette puller   Sutter Instruments Company Model P-30  
Microinjector   Narishige IM-300  
Pipette sharpener   Sutter Instruments Company Model BV-10  

References

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Cite This Article
Nijagal, A., Le, T., Wegorzewska, M., MacKenzie, T. C. A Mouse Model of in Utero Transplantation. J. Vis. Exp. (47), e2303, doi:10.3791/2303 (2011).

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