Name: a mouse model of in utero transplantation
Uploaded: 2011-01-27T17:00:00
Description: Watch this Scientific Journal Video about A Mouse Model of in Utero Transplantation at JoVE.com

我们要感谢我们的资金来源：再生医学临床研究员培训资助（AN）的加州理工学院，美国国家科学基金会（兆瓦），艾琳Perstein奖（TCM），美国外科医生学院（中医），美国小儿外科协会（中医），三月优生优育基金会（中医）。

1。制备注射移液器<ol><li>校准移液器拉马等玻璃吸管分离发生在15秒内（请参见制造商的说明，关于校准）。吸管将有锥度中分离出来。 </li><li>切断的吸管等，从开始锥度到距离吸管年底1.04厘米到1.05厘米年底。吸管的长度是成反比的吸管孔的口径。要知道，一个较长的吸管也处于弱势的提示，注射期间更容易破损。 </li><li>移液器的下一个步骤是创建一个光滑的斜面锐化冰山上的钻石磨刀轮。?...

<ol>
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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=in+utero+hematopoietic+stem+cell+transplantation:+ontogenic+opportunities+and+biologic+barriers.">in utero hematopoietic stem cell transplantation: ontogenic opportunities and biologic barriers.</a> Blood. 94, 2179-2191 (1999).</li><li>Merianos, 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=Maternal+alloantibodies+induce+a+postnatal+immune+response+that+limits+engraftment+following+in+utero+hematopoietic+cell+transplantation+in+mice.">Maternal alloantibodies induce a postnatal immune response that limits engraftment following in utero hematopoietic cell transplantation in mice.</a> J Clin Invest. 119, 2590-2600 (2009).</li><li>Peranteau, W. H., Endo, M., Adibe, O. O., Flake, A. 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F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+chimerism+threshold+predicts+sustained+engraftment+and+NK-cell+tolerance+in+prenatal+allogeneic+chimeras.">Early chimerism threshold predicts sustained engraftment and NK-cell tolerance in prenatal allogeneic chimeras.</a> Blood. 112, 5245-5253 (2008).</li><li>Mackenzie, T. C., Shaaban, A. F., Radu, A., Flake, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engraftment+of+bone+marrow+and+fetal+liver+cells+after+in+utero+transplantation+in+MDX+mice.">Engraftment of bone marrow and fetal liver cells after in utero transplantation in MDX mice.</a> J Pediatr Surg. 37, 1058-1064 (2002).</li><li>Hayashi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mixed+chimerism+following+in+utero+hematopoietic+stem+cell+transplantation+in+murine+models+of+hemoglobinopathy.">Mixed chimerism following in utero hematopoietic stem cell transplantation in murine models of hemoglobinopathy.</a> Exp Hematol. 31, 176-184 (2003).</li><li>Bouchard, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+transgene+expression+in+cardiac+and+skeletal+muscle+following+fetal+administration+of+adenoviral+or+adeno-associated+viral+vectors+in+mice.">Long-term transgene expression in cardiac and skeletal muscle following fetal administration of adenoviral or adeno-associated viral vectors in mice.</a> J Gene Med. 5, 941-950 (2003).</li><li>Meza, N. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rescue+of+pyruvate+kinase+deficiency+in+mice+by+gene+therapy+using+the+human+isoenzyme.">Rescue of pyruvate kinase deficiency in mice by gene therapy using the human isoenzyme.</a> Mol Ther. 17, 2000-2009 (2009).</li><li>MacKenzie, T. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+transduction+of+liver+and+muscle+after+in+utero+injection+of+lentiviral+vectors+with+different+pseudotypes.">Efficient transduction of liver and muscle after in utero injection of lentiviral vectors with different pseudotypes.</a> Mol Ther. 6, 349-358 (2002).</li><li>MacKenzie, T. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transduction+of+satellite+cells+after+prenatal+intramuscular+administration+of+lentiviral+vectors.">Transduction of satellite cells after prenatal intramuscular administration of lentiviral vectors.</a> J Gene Med. 7, 50-58 (2005).</li><li>Sabatino, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Persistent+expression+of+hF.IX+After+tolerance+induction+by+in+utero+or+neonatal+administration+of+AAV-1-F.IX+in+hemophilia+B+mice.">Persistent expression of hF.IX After tolerance induction by in utero or neonatal administration of AAV-1-F.IX in hemophilia B mice.</a> Mol Ther. 15, 1677-1685 (2007).</li><li>Mellor, A. L., Munn, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunology+at+the+maternal-fetal+interface:+lessons+for+T+cell+tolerance+and+suppression.">Immunology at the maternal-fetal interface: lessons for T cell tolerance and suppression.</a> Annu Rev Immunol. 18, 367-391 (2000).</li><li>Billingham, R. E., Brent, L., Medawar, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Actively+acquired+tolerance+of+foreign+cells.">Actively acquired tolerance of foreign cells.</a> Nature. 172, 603-606 (1953).</li><li>Endo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+transfer+to+ocular+stem+cells+by+early+gestational+intraamniotic+injection+of+lentiviral+vector.">Gene transfer to ocular stem cells by early gestational intraamniotic injection of lentiviral vector.</a> Mol Ther. 15, 579-587 (2007).</li><li>Waddington, S. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+transgene+expression+by+administration+of+a+lentivirus-based+vector+to+the+fetal+circulation+of+immuno-competent+mice.">Long-term transgene expression by administration of a lentivirus-based vector to the fetal circulation of immuno-competent mice.</a> Gene Ther. 10, 1234-1240 (2003).</li><li>Schachtner, S., Buck, C., Bergelson, J., Baldwin, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporally+regulated+expression+patterns+following+in+utero+adenovirus-mediated+gene+transfer.">Temporally regulated expression patterns following in utero adenovirus-mediated gene transfer.</a> Gene Ther. 6, 1249-1257 (1999).</li></ol>

比林汉姆，布伦特和梅达沃在50年前， 在子宫内移植在小鼠体内诱导外源蛋白的免疫耐受。从那时起，16几个变化，这种技术已被用于解决在免疫学和干细胞生物学的问题。 这里详细的协议是IUT的最方便的方法之一。胎肝提供了一个轻松的可视化的目标，并提供通过门户网站和肝静脉全身血液循环的访问。然而，一些已修改，以优化的时间和所需注射的位置。?...

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<td>Pipettes</td>
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<td>Kimble</td>
<td>71900-100</td>
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<td>Pipette puller</td>
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<td>Sutter Instruments Company</td>
<td>Model P-30</td>
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<td>Microinjector</td>
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<td>Narishige</td>
<td>IM-300</td>
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<td>Pipette sharpener</td>
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<td>Sutter Instruments Company</td>
<td>Model BV-10</td>
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a mouse model of in utero transplantation

在子宫内移植的干细胞和病毒，用于治疗人类胎儿先天性疾病的巨大潜力。例如， 在子宫内的造血干细胞移植（IUT）已成功地用于治疗重症联合免疫缺陷。1,2在其他几个条件的患者，然而，IUT的已经尝试没有成功。鉴于这些好坏参半的结果，可用性一个有效率的非​​人性化的模型来研究干细胞移植和基因治疗的生物后遗症，是推进这一领域的关键。我们和其他人使用IUT的研究在野生型小鼠4-7和遗传性疾病的影响因素在宫内移植造血干细胞成功植入小鼠模型。8,9胎儿环境也提供了相当大的优势在子宫内基因治疗的成功。例如，交付10的腺病毒，腺相关病毒，逆转录病毒11和慢病毒载体进入胎儿体内12,13 在子宫内导致在远离注射网站长期的基因表达的多器官的转导。因此，基因疗法可能被视为一个单基因疾病，如肌肉萎缩症或囊性纤维化的可能的治疗策略。 IUT的另一个潜在优势是能够到一个特定的抗原诱导免疫耐受。由于血友病因子IX的宽容早在发展成果引入到这种蛋白质的小鼠 。14 除了其使用在调查潜在的人类疗法，对IUT的小鼠模型可以是一个强大的工具，在发育和干细胞生物学研究的基本问题。例如，一个可以提供各种小分子诱导或抑制特定基因的表达在妊娠阶段的定义和操作的发展途径。这些改动的影响，可以在不同的时间点进行评估后，最初的移植。此外，还有一个可以移植到胎儿的环境，研究在非照射和泰然自若的主机环境中的干细胞分化的多能干或宗族的祖细胞。 在免疫学领域，发育和干细胞生物学内的IUT的小鼠模型已经提供了许多见解。在此视频为基础的协议，我们描述了一个步骤，循序渐进的方式执行IUT的小鼠胎儿，并概述这项技术的关键步骤和潜在的隐患。

Watch this Scientific Journal Video about A Mouse Model of in Utero Transplantation at JoVE.com

A Mouse Model of in Utero Transplantation

在小鼠模型<em&gt;在子宫内</em&gt;移植是一种多用途的工具，可用于胎儿干细胞移植和基因治疗研究的潜在的临床应用。在这个协议中，我们提出了一个执行此技术的一般方法

一个小鼠模型在子宫内移植

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus.  For example, in ...

Research

JoVE Journal

Medicine

A Mouse Model of in Utero Transplantation

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

肺动脉瓣使用异位心脏移植小鼠模型移植

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

科研

医学

Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

小鼠角膜移植模型，研究实体器官移植的最常见形式

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each ...

Transplantation Into the Mouse Ovarian Fat Pad

移植到小鼠卵巢脂肪垫

Orthotopic transplantation assays in mice are invaluable for studies of cell regeneration and neoplastic transformation. Common approaches for orthotopic ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

套筒技术在主动脉移植小鼠模型中的应用-教学视频

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique

应用改良袖套技术的小鼠胰腺移植模型

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the ...

Intravital Microscopy and Thrombus Induction in the Earlobe of a Hairless Mouse

活体显微镜及血栓诱导在无毛小鼠的耳垂

Thrombotic complications of vascular diseases are one leading cause of morbidity and mortality in industrial nations. Due to the complex interactions ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

小鼠体外循环模型: 一种新方法

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

Intraportal Transplantation of Pancreatic Islets in Mouse Model

胰岛门静脉移植小鼠模型的研究

Pancreatic islet transplantation to reduce hyperglycemia is highly successful in rodents with chemically-induced diabetes. The most common transplantation ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Veno-Venous Extracorporeal Membrane Oxygenation in a Mouse

小鼠静脉内膜氧化术

The use of extracorporeal membrane oxygenation (ECMO) has increased substantially in recent years. ECMO has become a reliable and effective therapy for ...