Name: fat-covered islet transplantation using epididymal white adipose tissue
Uploaded: 2021-05-25T14:00:00
Description: Watch this Scientific Journal Video about Fat-Covered Islet Transplantation Using Epididymal White Adipose Tissue at JoVE.com

这项研究由日本文部科学省的科学研究补助金（C）（19K09839，NS）资助。

以下过程分三个步骤执行。第一步包括在受体小鼠中诱导糖尿病和分离供体胰岛。第二步涉及移植前胰岛的准备。在第三步中，将胰岛移植到附睾脂肪组织上，并使用脂肪组织覆盖胰岛。之后，评估治疗效果。本研究中对小鼠的处理和实验程序符合“实验动物护理原则”（实验动物护理和使用指南，美国国立卫生研究院出版物第8版，2011年），实验方案获得福冈大学动物护理和使用委员会...

<ol>
	<li>Barton, F. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+in+outcomes+of+clinical+islet+transplantation:+1999-2010.">Improvement in outcomes of clinical islet transplantation: 1999-2010.</a> Diabetes Care. 35 (7), 1436-1445 (2012).</li><li>Balamurugan, A. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Islet+product+characteristics+and+factors+related+to+successful+human+islet+transplantation+from+the+Collaborative+Islet+Transplant+Registry+(CITR)+1999-2010.">Islet product characteristics and factors related to successful human islet transplantation from the Collaborative Islet Transplant Registry (CITR) 1999-2010.</a> American Journal of Transplantation. 14 (11), 2595-2606 (2014).</li><li>Collaborative Islet Transplant Registry. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Collaborative Islet Transplant Registry. Annual Report. , (2017).</li><li>Rajab, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Total+Pancreatectomy+and+Islet+Autotransplantation+Following+Treated+Hepatitis+C+Infection.">Total Pancreatectomy and Islet Autotransplantation Following Treated Hepatitis C Infection.</a> Cell Transplantation. 27 (10), 1569-1573 (2018).</li><li>Mellgren, A., Schnell Landstrom, A. H., Petersson, B., Andersson, 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+renal+subcapsular+site+offers+better+growth+conditions+for+transplanted+mouse+pancreatic+islet+cells+than+the+liver+or+spleen.">The renal subcapsular site offers better growth conditions for transplanted mouse pancreatic islet cells than the liver or spleen.</a> Diabetologia. 29 (9), 670-672 (1986).</li><li>Hiller, W. F., Klempnauer, J., Luck, R., Steiniger, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive+deterioration+of+endocrine+function+after+intraportal+but+not+kidney+subcapsular+rat+islet+transplantation.">Progressive deterioration of endocrine function after intraportal but not kidney subcapsular rat islet transplantation.</a> Diabetes. 40 (1), 134-140 (1991).</li><li>Yasunami, Y., Lacy, P. E., Finke, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+site+for+islet+transplantation--a+peritoneal-omental+pouch.">A new site for islet transplantation--a peritoneal-omental pouch.</a> Transplantation. 36 (2), 181-182 (1983).</li><li>Kin, T., Korbutt, G. S., Rajotte, R. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survival+and+metabolic+function+of+syngeneic+rat+islet+grafts+transplanted+in+the+omental+pouch.">Survival and metabolic function of syngeneic rat islet grafts transplanted in the omental pouch.</a> American Journal of Transplantation. 3 (3), 281-285 (2003).</li><li>Kasoju, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioengineering+a+pre-vascularized+pouch+for+subsequent+islet+transplantation+using+VEGF-loaded+polylactide+capsules.">Bioengineering a pre-vascularized pouch for subsequent islet transplantation using VEGF-loaded polylactide capsules.</a> Biomaterials Science. 8 (2), 631-647 (2020).</li><li>Sakata, N., Yoshimatsu, G., Kodama, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=White+Adipose+Tissue+as+a+Site+for+Islet+Transplantation.">White Adipose Tissue as a Site for Islet Transplantation.</a> Transplantology. 1 (2), 55-70 (2020).</li><li>Osama Gaber, A., Chamsuddin, A., Fraga, D., Fisher, J., Lo, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin+independence+achieved+using+the+transmesenteric+approach+to+the+portal+vein+for+islet+transplantation.">Insulin independence achieved using the transmesenteric approach to the portal vein for islet transplantation.</a> Transplantation. 77 (2), 309-311 (2004).</li><li>Fujita, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technique+of+endoscopic+biopsy+of+islet+allografts+transplanted+into+the+gastric+submucosal+space+in+pigs.">Technique of endoscopic biopsy of islet allografts transplanted into the gastric submucosal space in pigs.</a> Cell Transplantation. 22 (12), 2335-2344 (2013).</li><li>Sakata, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strategy+for+clinical+setting+in+intramuscular+and+subcutaneous+islet+transplantation.">Strategy for clinical setting in intramuscular and subcutaneous islet transplantation.</a> Diabetes/Metabolism Research and Reviews. 30 (1), 1-10 (2014).</li><li>Cantarelli, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplant+Site+Influences+the+Immune+Response+After+Islet+Transplantation:+Bone+Marrow+Versus+Liver.">Transplant Site Influences the Immune Response After Islet Transplantation: Bone Marrow Versus Liver.</a> Transplantation. 101 (5), 1046-1055 (2017).</li><li>White, S. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+risks+of+total+pancreatectomy+and+splenic+islet+autotransplantation.">The risks of total pancreatectomy and splenic islet autotransplantation.</a> Cell Transplantation. 9 (1), 19-24 (2000).</li><li>Itoh, T., Nishinakamura, H., Kumano, K., Takahashi, H., Kodama, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Spleen+Is+an+Ideal+Site+for+Inducing+Transplanted+Islet+Graft+Expansion+in+Mice.">The Spleen Is an Ideal Site for Inducing Transplanted Islet Graft Expansion in Mice.</a> PLoS One. 12 (1), 0170899 (2017).</li><li>Sakata, N., Yoshimatsu, G., Kodama, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Spleen+as+an+Optimal+Site+for+Islet+Transplantation+and+a+Source+of+Mesenchymal+Stem+Cells.">The Spleen as an Optimal Site for Islet Transplantation and a Source of Mesenchymal Stem Cells.</a> International Journal of Molecular Sciences. 19 (5), (2018).</li><li>Sakata, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+rat-to-mouse+bioartificial+pancreas+xenotransplantation+on+diabetic+renal+damage+and+survival.">Effect of rat-to-mouse bioartificial pancreas xenotransplantation on diabetic renal damage and survival.</a> Pancreas. 32 (3), 249-257 (2006).</li><li>Nagaya, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+bioengineered+islet+cell+sheets+for+the+treatment+of+diabetes+mellitus.">Effectiveness of bioengineered islet cell sheets for the treatment of diabetes mellitus.</a> Journal of Surgical Research. 227, 119-129 (2018).</li><li>Weaver, J. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vasculogenic+hydrogel+enhances+islet+survival,+engraftment,+and+function+in+leading+extrahepatic+sites.">Vasculogenic hydrogel enhances islet survival, engraftment, and function in leading extrahepatic sites.</a> Science Advances. 3 (6), 1700184 (2017).</li><li>Dufour, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+ectopic+site+for+islet+transplantation,+using+biodegradable+scaffolds.">Development of an ectopic site for islet transplantation, using biodegradable scaffolds.</a> Tissue Engineering. 11 (9-10), 1323-1331 (2005).</li><li>Chen, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+epididymal+fat+pad+as+a+transplant+site+for+minimal+islet+mass.">The epididymal fat pad as a transplant site for minimal islet mass.</a> Transplantation. 84 (1), 122-125 (2007).</li><li>Sakata, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+Transplanted+Islet+Engraftment+in+Visceral+White+Adipose+Tissue.">Mechanism of Transplanted Islet Engraftment in Visceral White Adipose Tissue.</a> Transplantation. 104 (12), 2516-2527 (2020).</li><li>Navarro-Requena, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PEG+hydrogel+containing+calcium-releasing+particles+and+mesenchymal+stromal+cells+promote+vessel+maturation.">PEG hydrogel containing calcium-releasing particles and mesenchymal stromal cells promote vessel maturation.</a> Acta Biomaterialia. 67, 53-65 (2018).</li><li>Phelps, E. A., Headen, D. M., Taylor, W. R., Thule, P. M., Garcia, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vasculogenic+bio-synthetic+hydrogel+for+enhancement+of+pancreatic+islet+engraftment+and+function+in+type+1+diabetes.">Vasculogenic bio-synthetic hydrogel for enhancement of pancreatic islet engraftment and function in type 1 diabetes.</a> Biomaterials. 34 (19), 4602-4611 (2013).</li><li>Manzoli, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunoisolation+of+murine+islet+allografts+in+vascularized+sites+through+conformal+coating+with+polyethylene+glycol.">Immunoisolation of murine islet allografts in vascularized sites through conformal coating with polyethylene glycol.</a> American Journal of Transplantation. 18 (3), 590-603 (2018).</li><li>Gotoh, M., Maki, T., Kiyoizumi, T., Satomi, S., Monaco, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+method+for+isolation+of+mouse+pancreatic+islets.">An improved method for isolation of mouse pancreatic islets.</a> Transplantation. 40 (4), 437-438 (1985).</li><li>Brandhorst, D., Brandhorst, H., Hering, B. J., Bretzel, R. G. <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+survival,+morphology+and+in+vitro+function+of+isolated+pig+islets+under+different+culture+conditions.">Long-term survival, morphology and in vitro function of isolated pig islets under different culture conditions.</a> Transplantation. 67 (12), 1533-1541 (1999).</li><li>Noguchi, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-temperature+preservation+of+isolated+islets+is+superior+to+conventional+islet+culture+before+islet+transplantation.">Low-temperature preservation of isolated islets is superior to conventional islet culture before islet transplantation.</a> Transplantation. 89 (1), 47-54 (2010).</li><li>Itoh, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low+temperature+condition+prevents+hypoxia-induced+islet+cell+damage+and+HMGB1+release+in+a+mouse+model.">Low temperature condition prevents hypoxia-induced islet cell damage and HMGB1 release in a mouse model.</a> Cell Transplantation. 21 (7), 1361-1370 (2012).</li><li>Komatsu, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimizing+Temperature+and+Oxygen+Supports+Long-term+Culture+of+Human+Islets.">Optimizing Temperature and Oxygen Supports Long-term Culture of Human Islets.</a> Transplantation. 103 (2), 299-306 (2019).</li><li>Unger, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lipid+overload+and+overflow:+metabolic+trauma+and+the+metabolic+syndrome.">Lipid overload and overflow: metabolic trauma and the metabolic syndrome.</a> Trends in Endocrinology, Metabolism. 14 (9), 398-403 (2003).</li><li>Mao, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+macroporous+heparin-releasing+silk+fibroin+scaffold+improves+islet+transplantation+outcome+by+promoting+islet+revascularisation+and+survival.">A macroporous heparin-releasing silk fibroin scaffold improves islet transplantation outcome by promoting islet revascularisation and survival.</a> Acta Biomaterialia. 59, 210-220 (2017).</li><li>Wang, K., Wang, X., Han, C. S., Chen, L. 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脂肪覆盖的胰岛移植方法结合了两种不同移植技术的技术：腹膜内胰岛移植和脂肪内组织胰岛移植。由于附睾白色脂肪组织的表面膜被认为是被腹膜覆盖并附着在附睾上的白色脂肪组织，因此脂肪覆盖胰岛移植方法在解剖学上可归类为腹膜内胰岛移植的一种。然而，将胰岛输送给受体动物的技术更类似于脂肪内组织胰岛移植中使用的技术。我们的数据显示，脂肪覆盖胰岛移植方法的治疗效果优于?...

胰岛移植是重度糖尿病患者的细胞替代疗法。最近的报告显示，移植后三年的胰岛素独立率提高到44%1 ，在接受超过600，000个胰岛当量的接受者中，约有80%实现了胰岛素独立2。此外，在最新的协作胰岛移植登记报告中，显示超过70%的单独胰岛移植患者的空腹血糖水平在5年内保持在60-140mg / dL。该研究还确定，在肾移植后单独接受胰岛移植或胰岛移植的患者中，约有90%在5年内没有发生任何严重的低血糖事件3。
尽管这种治疗的临床结果一直在改善，但仍必须解决一些局限性，包括建立最佳移植部位的必要性。肝脏是临床胰岛移植的典型移植部位，因为它是可容纳大量胰岛的最大器官。然而，在一些患者中，肝脏不可用（例如，由于门静脉高压、肝炎和/或肝硬化4），因此其他部位，包括肾囊下间隙5，6、网膜袋7，8，9，10、肠系膜 11、胃肠道 12、骨骼肌 13、皮下组织 13、骨髓 14 和脾脏15，16，17，已被考虑作为替代移植部位。
虽然腹腔内胰岛移植在局部麻醉下容易进行，使腹腔内腔成为临床胰岛移植的吸引人部位，但在移植时，胰岛分散在整个腹腔内，难以进行胰岛植入检测和成功植入确认。因此，腹腔内尚未被广泛认为是理想的临床移植部位。相反，它经常被用作临床前研究的对照模型，以研究移植封装18 和生物工程胰岛19的有效性。然而，由于在进行准确的植入评估方面存在挑战，很难在生物工程胰岛和对照胰岛之间进行精确比较。
相比之下，在网膜袋8、肠系膜和其他肝外部位使用腹膜内白色脂肪组织已被充分报道10，20，21，22，23，许多调查使用白色脂肪组织移植的生物工程胰岛功能的研究能够报告有希望的治疗结果20，24，25，26.由于附睾脂肪组织的使用有助于移植胰岛的检测，因此开发了利用附睾脂肪组织的“脂肪覆盖胰岛移植方法”，以克服腹腔内胰岛移植的局限性。本文描述了使用附睾脂肪组织进行脂肪覆盖的胰岛移植。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>4-0 Nylon</td>
			<td>Alfresa</td>
			<td>ER2004NA45-KF2</td>
			<td>Closing abdomen</td>
		</tr>
		<tr>
			<td>Alexa 488-conjugated donkey anti-guinea pig</td>
			<td>Jackson Immunoresearch</td>
			<td>706-546-148</td>
			<td>Secondary antibody for insulin antibody</td>
		</tr>
		<tr>
			<td>Alexa 647-conjugated donkey anti-rabbit</td>
			<td>Jackson Immunoresearch</td>
			<td>711-606-152</td>
			<td>Secondary antibody for von Willebrand factor antibody</td>
		</tr>
		<tr>
			<td>DMEM, low glucose, pyruvate</td>
			<td>ThermoFisher Scientific</td>
			<td>11885084</td>
			<td>Culturing islets, transplanting islets</td>
		</tr>
		<tr>
			<td>Eosin</td>
			<td>Fujifilm Wako Chemicals</td>
			<td>051-06515</td>
			<td>Using for staining tissue by eosin</td>
		</tr>
		<tr>
			<td>Eppendorf Safe-Lock Tubes, 1.5 mL</td>
			<td>Eppendorf</td>
			<td>30120086</td>
			<td>Collecting islets&#160;</td>
		</tr>
		<tr>
			<td>Falcon 15 mL Conical Centrifuge Tubes</td>
			<td>Corning</td>
			<td>352095</td>
			<td>Collecting islets</td>
		</tr>
		<tr>
			<td>Falcon 40 &#181;m Cell Strainer</td>
			<td>Falcon</td>
			<td>352340</td>
			<td>Using for separating islets from other pancreatic tissue</td>
		</tr>
		<tr>
			<td>Falcon 50 mL Conical Centrifuge Tubes</td>
			<td>Corning</td>
			<td>352070</td>
			<td>Discarding excessive medium/buffer</td>
		</tr>
		<tr>
			<td>Guinea pig anti-insulin</td>
			<td>Agilent Technologies Japan, Ltd. (Dako)</td>
			<td>IR002</td>
			<td>Primary antibody for murine insulin</td>
		</tr>
		<tr>
			<td>Hematoxylin</td>
			<td>Muto Pure Chemicals Co., Ltd.</td>
			<td>30002</td>
			<td>Using for staining tissue by hematoxylin</td>
		</tr>
		<tr>
			<td>Isodine solution 10%</td>
			<td>Shionogi&#38;Co., Ltd.</td>
			<td>no catalog number</td>
			<td>Using for disinfection</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Fujifilm Wako Chemicals</td>
			<td>095-06573</td>
			<td>Using for anesthesia</td>
		</tr>
		<tr>
			<td>Labcon 1000 &#181;L ZapSilk Low Retention Pipette Tips</td>
			<td>Labcon</td>
			<td>1177-965-008</td>
			<td>Using for separating islets from other pancreatic tissue</td>
		</tr>
		<tr>
			<td>Labcon 200 &#181;L ZapSilk Low Retention Pipette Tips</td>
			<td>Labcon</td>
			<td>1179-965-008</td>
			<td>Using for seeding islets onto epididymal white adipose tissue</td>
		</tr>
		<tr>
			<td>Mintsensor</td>
			<td>Sanwa Kagaku Kenkyusho Co. Ltd.,</td>
			<td>8AEB02E</td>
			<td>Using for monitoring blood glucose</td>
		</tr>
		<tr>
			<td>Pipetteman P-1000</td>
			<td>Gilson</td>
			<td>F123602</td>
			<td>Using for separating islets from other pancreatic tissue</td>
		</tr>
		<tr>
			<td>Pipetteman P-200</td>
			<td>Gilson</td>
			<td>F123601</td>
			<td>Using for seeding islets onto epididymal white adipose tissue</td>
		</tr>
		<tr>
			<td>Rabbit anti-vWF</td>
			<td>Abcam</td>
			<td>ab6994</td>
			<td>Primary antibody for murine von Willebrand factor</td>
		</tr>
	</tbody>
</table>

fat-covered islet transplantation using epididymal white adipose tissue

胰岛移植是严重糖尿病的细胞替代疗法。腹腔通常是该手术的移植部位。然而，腹腔内胰岛移植存在一定的局限性，包括移植效果差，移植物检测能力困难，移植后分析缺乏移植切除能力。本文采用“脂肪覆盖胰岛移植”，一种利用附睾白色脂肪组织的腹膜内胰岛移植方法，用于评估生物工程胰岛的治疗效果。该方法的简单之处在于将胰岛播种到附睾白色脂肪组织上，并利用组织覆盖胰岛。虽然这种方法可以归类为腹膜内胰岛移植技术，但它与脂肪内组织胰岛移植具有共同特征。然而，脂肪覆盖的胰岛移植方法显示出比脂肪内组织胰岛移植更强大的治疗效果，包括改善血糖和血浆胰岛素水平以及移植物切除的潜力。我们建议采用这种方法来评估胰岛植入白色脂肪组织的机制和生物工程胰岛的治疗效果。

为了比较脂肪覆盖胰岛移植与腹膜内胰岛移植后的移植效果，将相同数量的胰岛植入对照受体糖尿病动物左侧结肠旁间隙的腹膜上。与腹膜内胰岛移植小鼠相比，观察到脂肪覆盖胰岛移植小鼠的血糖水平逐渐显着降低（p = 0.0023; 图3A）。移植后1个月，脂肪覆盖胰岛移植小鼠的血糖维持在低于腹膜内胰岛移植小鼠的血糖水平，通过腹膜内葡萄糖耐量试验评估（p = 0.0046; <strong cl...

Watch this Scientific Journal Video about Fat-Covered Islet Transplantation Using Epididymal White Adipose Tissue at JoVE.com

Fat-Covered Islet Transplantation Using Epididymal White Adipose Tissue

这种脂肪覆盖的胰岛移植方法适用于检测腹膜腔内的移植胰岛。值得注意的是，它不需要使用生物结合剂或缝合。

使用附睾白色脂肪组织进行脂肪覆盖的胰岛移植

Islet transplantation is a cellular replacement therapy for severe diabetes mellitus. The intraperitoneal cavity is typically the transplant site for this ...

该协议可以评估胰岛植入白色脂肪组织的机制。这种脂肪覆盖的胰岛移植方法的主要优点是它在技术上易于复制。该方法可应用于其他疾病模型。例如，它可用于创建癌症的小鼠模型。演示该程序的将是福冈大学副教授Naoaki Sakata。在解剖显微镜下，使用P 200微量移液管从胰腺消化中去除任何额外的胰岛成分，包括腺泡和纤维组织。然后使用细胞过滤器过滤掉单个腺泡细胞。将过滤后的胰岛转移到含有补充有牛血清或白蛋白的适当培养基或缓冲溶液的新培养皿中，并旋转培养皿以将胰岛定位在中心。使用 P 200 微量移液器，将各个胰岛挑入适当的收集管中。将一个新的 40 微米细胞过滤器放在 50 毫升塑料管的顶部。使用1000微升移液器将胰岛添加到过滤器中，以分离胰岛和单个腺泡细胞。使用镊子将过滤器倒置在新的60或100毫升大小的未处理培养皿上，该培养皿含有培养基或适当的缓冲溶液，并补充有牛血清或白蛋白。使用新鲜培养基或缓冲液将胰岛冲洗到新的培养皿中，然后在培养皿中加入足够的培养基或缓冲液，以达到约20毫升的总体积。在显微镜下计数胰岛，并根据供体动物的数量在单独的1.5毫升塑料离心管之间平均分配胰岛的数量。在室温下以2， 100G离心胰岛一分钟，弃去上清液。大约 20 至 30 微升的残留溶液通常会留在管中。从腹部去除毛发以防止感染后，将鼠标置于仰卧位。对腹部和腹股沟区域进行消毒后，在较低的中位区域切开皮肤，形成长约2厘米的皮肤切口。用镊子或牵开器夹住左腹壁，并将组织拉到小鼠的左侧以固定手术区域。使用棉签将小肠和大肠移动到小鼠的右侧。腹腔中的左表皮白色脂肪组织位于左腹股沟区域。将表皮白色脂肪组织和左侧睾丸移动到腹部外侧并伸展组织。使用 P 200 微量移液器从一个 1.5 毫升管中收集整个胰岛体积，注意收集时管中没有胰岛。让收集的胰岛通过重力沉淀在移液器的尖端。将微量移液器吸头轻轻放在膨胀的脂肪组织上，注意防止吸头中的培养基或缓冲液过度冲洗。小心地将胰岛播种到组织上。播种后，在解剖显微镜下确认胰岛的正确放置。用表皮白色脂肪组织覆盖胰岛，然后将左睾丸置于游离体白色脂肪组织下，并将组织返回腹膜腔。使用4-o缝合线将皮肤封闭成两层。完成后，将鼠标放在加热灯下并监视它，直到完全恢复。为了比较脂肪覆盖胰岛移植与腹膜内胰岛移植后的移植效果，将相同数量的胰岛植入对照受体糖尿病动物左侧结肠旁间隙的腹膜上。观察到脂肪覆盖胰岛移植小鼠的血糖水平与腹膜内胰岛移植小鼠相比逐渐显着降低。移植后一个月进行腹膜内葡萄糖耐量试验。脂肪覆盖胰岛移植小鼠的血糖维持在低于腹膜内胰岛移植小鼠观察到的水平。组织学检查用于评估胰岛植入表皮白色脂肪组织的情况。在脂肪覆盖的胰岛移植受者动物中，苏木精始新世染色显示表皮白色脂肪组织中存在胰岛。此外，胰岛素阳性胰岛的荧光偶联抗体染色有助于检测所有受体小鼠表皮白色脂肪组织中的血管性血友病因子阳性微血管。为了成功进行此过程，请确保让胰岛完全沉降在移液器的尖端，并将它们播种到脂肪组织上而不会溢出。

Research

JoVE Journal

Medicine

A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

小鼠胰岛的分离和包膜下肾移植的方法

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize ...

科研

医学

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 ...

Cerenkov Luminescence Imaging of Interscapular Brown Adipose Tissue

肩胛间褐色脂肪组织的切伦科夫发光成像

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

封装产热前脂肪细胞移植到脂肪组织仓库

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

Creation and Transplantation of an Adipose-derived Stem Cell (ASC) Sheet in a Diabetic Wound-healing Model

Creation and Transplantation of an Adipose-derived Stem Cell ASC Sheet in a Diabetic Wound-healing Model

创作和移植的糖尿病伤口愈合模型中脂肪源性干细胞 (ASC) 表

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

应用 Videomicroscopy 评价人脂肪组织微血管功能

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

脂肪组织干细胞移植对大鼠部分结肠切除术后渗漏的减少

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Characterization of Immune Cells in Human Adipose Tissue by Using Flow Cytometry

应用流式细胞仪对人体脂肪组织免疫细胞的表征

Infiltration of immune cells in the subcutaneous and visceral adipose tissue (AT) deposits leads to a low-grade inflammation contributing to the ...

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

18F-fdg Petct 对活性人棕色脂肪组织的全身和区域定量

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets

Inguinal Subcutaneous White Adipose Tissue ISWAT Transplantation Model of Murine Islets

毛里岛皮下白脂肪组织（ISWAT）移植模型

Pancreatic islet transplantation is a well-established therapeutic treatment for type 1 diabetes. The kidney capsule is the most commonly used site for ...