Name: collection protocol for human pancreas
Uploaded: 2012-05-23T12:00:00
Description: Watch this Scientific Journal Video about Collection Protocol for Human Pancreas at JoVE.com

लेखकों &#39;दाताओं के परिवारों और अंग प्रत्यारोपण खरीद में शामिल उनके विशेषज्ञ की सहायता के लिए इस शोध और मारिया मार्टिनो और इरीना Kusmartseva के संगठन धन्यवाद. यह काम अग्नाशय के मधुमेह के साथ अंग दाताओं के लिए नेटवर्क के समर्थन में किशोर मधुमेह अनुसंधान फाउंडेशन (MC-T., एमए) द्वारा वित्त पोषित किया गया था.

1. सेट अप <ol><li> एक पेंसिल के साथ, या स्वतः एक कैसेट प्रिंटर के साथ में तेल ब्लॉक मैन्युअल के लिए लेबल कैसेट. दाता पहचानकर्ता (CaseID), नमूना प्रकार, अशेष भाजक संख्या, और किसी भी अतिरिक्त अद्वितीय पहचान (प्रकरण, का?...

<ol>
	<li>Brissova, M. <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+human+pancreatic+islet+architecture+and+composition+by+laser+scanning+confocal+microscopy.">Assessment of human pancreatic islet architecture and composition by laser scanning confocal microscopy.</a> J. Histochem. Cytochem. 53, 1087-1097 (2005).</li><li>Rahier, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pancreatic+polypeptide+cells+in+the+human+pancreas:+the+effects+of+age+and+diabetes.">The pancreatic polypeptide cells in the human pancreas: the effects of age and diabetes.</a> J. Clin. Endocrinol. Metab. 56, 441-444 (1983).</li><li>Uchida, T., Takada, T., Ammori, B. J., Suda, K., Takahashi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+reconstruction+of+the+ventral+and+dorsal+pancreas:+a+new+insight+into+anatomy+and+embryonic+development.">Three-dimensional reconstruction of the ventral and dorsal pancreas: a new insight into anatomy and embryonic development.</a> J. Hepatobiliary Pancreat. Surg. 6, 176-180 (1999).</li><li>Wittingen, J., Frey, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Islet+concentration+in+the+head,+body,+tail+and+uncinate+process+of+the+pancreas.">Islet concentration in the head, body, tail and uncinate process of the pancreas.</a> Ann. Surg. 179, 412-414 (1974).</li><li>Rahier, J., Goebbels, R., Henquin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+composition+of+the+human+diabetic+pancreas.">Cellular composition of the human diabetic pancreas.</a> Diabetologia. 24, 366-371 (1983).</li><li>Vaught, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+NCI+perspective+on+creating+sustainable+biospecimen+resources.">An NCI perspective on creating sustainable biospecimen resources.</a> J. Natl. Cancer Inst. Monogr. 42, 1-7 (2011).</li><li>Gianani, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Initial+results+of+screening+of+nondiabetic+organ+donors+for+expression+of+islet+autoantibodies.">Initial results of screening of nondiabetic organ donors for expression of islet autoantibodies.</a> J. Clin. Endocrinol. Metab. 91, 1855-1861 (2006).</li><li>Tauriainen, S., Salmela, K., Rantala, I., Knip, M., Hy&#246;ty, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collecting+high-quality+pancreatic+tissue+for+experimental+study+from+organ+donors+with+signs+of+&#946;-cell+autoimmunity.">Collecting high-quality pancreatic tissue for experimental study from organ donors with signs of &#946;-cell autoimmunity.</a> Diabetes Metab. Res. Rev. 26, 585-592 (2010).</li><li>In't Veld, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Screening+for+insulitis+in+adult+autoantibody-positive+organ+donors.">Screening for insulitis in adult autoantibody-positive organ donors.</a> Diabetes. 56, 2400-2404 (2007).</li><li>T&#246;ns, H. A., Terpstra, O. T., Bouwman, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneity+of+human+pancreata+in+perspective+of+the+isolation+of+the+islets+of+langerhans.">Heterogeneity of human pancreata in perspective of the isolation of the islets of langerhans.</a> Transplant Proc. 40, 367-369 (2008).</li></ol>

इस प्रक्रिया के लक्ष्य के लिए एक मानक अग्न्याशय प्रसंस्करण है कि कई संग्रह साइटों के पार संगत हो सकता है परिभाषित करने और इस तरह अलग अलग समूहों द्वारा प्राप्त परिणामों की तुलना की अनुमति है. विच्छेदन व...

<table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td><td> टिप्पणियाँ </td></tr><tr><td> 22 ब्लेड के साथ नलियां सहित विच्छेदन के यंत्र </td><td> विभिन्न </td><td> विभिन्न </td><td></td></tr><tr><td> अपकेंद्रित्र ट्यूबों (15 और 50 मिलीलीटर) </td><td> फिशर साइंटिफिक </td><td> 430,828 05-539-5 </td><td> बाँझ </td></tr><tr><td> Cryotubes (1.8 मिलीग्राम) </td><td> VWR उत्तर अमेरिका </td><td> 89004-300 </td><td> बाँझ, बार कोड वैकल्पिक </td></tr><tr><td> RNAlater </td><td> Ambion </td><td> AM7021 </td><td></td></tr><tr><td> सर्जिकल पैथोलॉजी स्याही </td><td></td><td></td><td></td></tr><tr><td> कैसेट </td><td> मर्सिडीज मेडिकल </td><td> कैस ECO109 </td><td></td></tr><tr><td> 10 तटस्थ% Formalin के बफर </td><td> फिशर साइंटिफिक </td><td> 23-245-685 </td><td></td></tr><tr><td> अक्टूबर बर्फ़ीली मीडिया </td><td> TissueTek </td><td> 4583 </td><td></td></tr><tr><td> Molds </td><td> ThermoScientific </td><td> 58,952 </td><td> विभिन्न आकारों </td></tr><tr><td> Isopentane </td><td> फिशर साइंटिफिक </td><td> : O3551 - 4 </td><td> अक्टूबर द्रुतशीतन स्नान </td></tr><tr><td> सूखी बर्फ </td><td></td><td></td><td> अक्टूबर द्रुतशीतन स्नान </td></tr><tr><td> देवर कुप्पी में तरल नाइट्रोजन </td><td></td><td></td><td> ठंड cryovials स्नैप </td></tr><tr><td> सेल संस्कृति डिश (100mm) </td><td> कोर्निंग </td><td> 430,167 </td><td> बाँझ </td></tr><tr><td> Hyclone DMEF </td><td> ThermoScientific </td><td> SH30023.01 </td><td> बाँझ दुकान, प्रशीतित, glutamine शामिल </td></tr><tr> &lt;&gt; भ्रूण गोजातीय सीरम td </td><td> Cellgro </td><td> 35-010-CV </td><td> बाँझ, दुकान जमी aliquots </td></tr><tr><td> पेनिसिलिन स्ट्रेप्टोमाइसिन / / ऐंटिफंगल </td><td> GIBCO </td><td> 15240-062 </td><td> बाँझ, दुकान जमी aliquots </td></tr><tr><td> 9 आयल लिखें </td><td> रिचर्ड - एलन वैज्ञानिक </td><td> 8337 </td><td></td></tr><tr><td> डी - पीबीएस 1X </td><td> GIBCO </td><td> 14190-144 </td><td> बाँझ </td></tr><tr><td> 16% Paraformaldehyde </td><td> EM विज्ञान </td><td> 15,710 </td><td> ताजा दुकान, प्रशीतित </td></tr><tr><td> 8% Glutaraldehyde के </td><td> EM विज्ञान </td><td> 16,019 </td><td> ताजा दुकान, प्रशीतित </td></tr><tr><td> जैवसुरक्षा स्तर 2 हुड </td><td> विभिन्न </td><td></td><td></td></tr><tr><td> अल्ट्रा कम फ्रीजर </td><td> विभिन्न </td><td></td><td></td></tr><tr><td> Sakura कैसेट्स प्रिंटर </td><td> Sakura </td><td></td><td></td></tr><tr><td> Sakura VIP300 </td><td> Sakura </td><td></td><td> स्वचालित आयल प्रोसेसर </td></tr><tr><td> पैराफिन एम्बेडिंग स्टेशन </td><td> ThermoFisher </td><td></td><td></td></tr></tbody></table> तालिका 1 विशिष्ट अभिकर्मकों और उपकरण.

collection protocol for human pancreas

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation of pancreas recovered from cadaveric organ donors. The pancreas is divided into 3 main regions (head, body, tail) followed by serial transverse sections throughout the medial to lateral axis. Alternating sections are used for fixed paraffin and fresh frozen blocks and remnant samples are minced for snap frozen sample preparations, either with or without RNAse inhibitors, for DNA, RNA, or protein isolation. The overall goal of the pancreas dissection procedure is to sample the entire pancreas while maintaining anatomical orientation.
Endocrine cell heterogeneity in terms of islet composition, size, and numbers is reported for human islets compared to rodent islets 1. The majority of human islets from the pancreas head, body and tail regions are composed of insulin-containing &beta;-cells followed by lower proportions of glucagon-containing &alpha;-cells and somatostatin-containing &delta;-cells. Pancreatic polypeptide-containing PP cells and ghrelin-containing epsilon cells are also present but in small numbers. In contrast, the uncinate region contains islets that are primarily composed of pancreatic polypeptide-containing PP cells 2. These regional islet variations arise from developmental differences. The pancreas develops from the ventral and dorsal pancreatic buds in the foregut and after rotation of the stomach and duodenum, the ventral lobe moves and fuses with the dorsal 3. The ventral lobe forms the posterior portion of the head including the uncinate process while the dorsal lobe gives rise to the rest of the organ. Regional pancreatic variation is also reported with the tail region having higher islet density compared to other regions and the dorsal lobe-derived components undergoing selective atrophy in type 1 diabetes4,5.
Additional organs and tissues are often recovered from the organ donors and include pancreatic lymph nodes, spleen and non-pancreatic lymph nodes. These samples are recovered with similar formats as for the pancreas with the addition of isolation of cryopreserved cells. When the proximal duodenum is included with the pancreas, duodenal mucosa may be collected for paraffin and frozen blocks and minced snap frozen preparations.

Watch this Scientific Journal Video about Collection Protocol for Human Pancreas at JoVE.com

Collection Protocol for Human Pancreas

इस वीडियो एकाधिक भंडारण स्वरूपों में मानव अग्न्याशय के प्रसंस्करण के लिए एक विच्छेदन प्रक्रिया को दर्शाता है. क्षेत्रीय आइलेट संरचना और घनत्व की परिभाषा की अनुमति संरचनात्मक अभिविन्यास अग्नाशय क्षेत्रों भर में बनाए रखा है.

मानव अग्नाशय के लिए संग्रह प्रोटोकॉल

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation ...

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