Name: intrasplenic transplantation of hepatocytes after partial hepatectomy in nod.scid mice
Uploaded: 2018-02-10T13:00:00
Description: Watch this Scientific Journal Video about Intrasplenic Transplantation of Hepatocytes After Partial Hepatectomy in NOD.SCID Mice at JoVE.com

Dette arbejde blev støttet af core tilskuddet modtaget fra Institut for bioteknologi, Indiens regering til de nationale Institut for Immunology, New Delhi. Dr. Bhattacharjee nuværende adresse er afdelingen for gastroenterologi, hepatologi og ernæring, children's Hospital Los Angeles.

Procedurerne i denne protokol er blevet godkendt af den institutionelle dyr etiske komité i den nationale Institut for Immunology, New Delhi. Seriel referencenummeret for godkendelsen er IAEC #319/13.
Bemærk: Der er gode ressourcer på almen kirurgi procedurer20 og specifikke protokoller for gnavere kirurgi21. For dem, der gør dyrenes kirurgi for første gang, er det anbefales til omfattende praksis kirurgiske procedurer på dummies f...

<ol>
	<li>Nussler, 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=Present+status+and+perspectives+of+cell-based+therapies+for+liver+diseases.">Present status and perspectives of cell-based therapies for liver diseases.</a> J. Hepatol. 45 (1), 144-159 (2006).</li><li>Ponder, K. P., 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=Mouse+hepatocytes+migrate+to+liver+parenchyma+and+function+indefinitely+after+intrasplenic+transplantation.">Mouse hepatocytes migrate to liver parenchyma and function indefinitely after intrasplenic transplantation.</a> Proc. Natl. Acad. Sci. U.S.A. 88 (4), 1217-1221 (1991).</li><li>Kokudo, N., Horimoto, H., Ishida, K., Takahashi, S., Nozawa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Allogeneic+hepatocyte+and+fetal+liver+transplantation+and+xenogeneic+hepatocyte+transplantation+for+Nagase's+analbuminemic+rats.">Allogeneic hepatocyte and fetal liver transplantation and xenogeneic hepatocyte transplantation for Nagase's analbuminemic rats.</a> Cell Transplant. 5 (5 Suppl 1), S21-S22 (1996).</li><li>Christ, B., Bruckner, S., Stock, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatic+transplantation+of+mesenchymal+stem+cells+in+rodent+animal+models.">Hepatic transplantation of mesenchymal stem cells in rodent animal models.</a> Methods Mol. Biol. 698, 315-330 (2011).</li><li>Fox, I. J., Roy-Chowdhury, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatocyte+transplantation.">Hepatocyte transplantation.</a> J. Hepatol. 40 (6), 878-886 (2004).</li><li>Glanemann, 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=Transplantation+of+monocyte-derived+hepatocyte-like+cells+(NeoHeps)+improves+survival+in+a+model+of+acute+liver+failure.">Transplantation of monocyte-derived hepatocyte-like cells (NeoHeps) improves survival in a model of acute liver failure.</a> Ann. Surg. 249 (1), 149-154 (2009).</li><li>Rahman, T. M., Hodgson, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+acute+hepatic+failure.">Animal models of acute hepatic failure.</a> Int. J. Exp. Pathol. 81 (2), 145-157 (2000).</li><li>Zhang, L., 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=Granulocyte+colony-stimulating+factor+treatment+ameliorates+liver+injury+and+improves+survival+in+rats+with+D-galactosamine-induced+acute+liver+failure.">Granulocyte colony-stimulating factor treatment ameliorates liver injury and improves survival in rats with D-galactosamine-induced acute liver failure.</a> Toxicol. Lett. 204 (1), 92-99 (2011).</li><li>Gardner, C. R., 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=Role+of+nitric+oxide+in+acetaminophen-induced+hepatotoxicity+in+the+rat.">Role of nitric oxide in acetaminophen-induced hepatotoxicity in the rat.</a> Hepatology. 27 (3), 748-754 (1998).</li><li>Nardo, 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=Successful+treatment+of+CCL4-induced+acute+liver+failure+with+portal+vein+arterialization+in+the+rat.">Successful treatment of CCL4-induced acute liver failure with portal vein arterialization in the rat.</a> Transplant Proc. 38 (4), 1187-1189 (2006).</li><li>Sathyasaikumar, K. 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=Fulminant+hepatic+failure+in+rats+induces+oxidative+stress+differentially+in+cerebral+cortex,+cerebellum+and+pons+medulla.">Fulminant hepatic failure in rats induces oxidative stress differentially in cerebral cortex, cerebellum and pons medulla.</a> Neurochem. Res. 32 (3), 517-524 (2007).</li><li>Wu, J., 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=Laennec+protects+murine+from+concanavalin+A-induced+liver+injury+through+inhibition+of+inflammatory+reactions+and+hepatocyte+apoptosis.">Laennec protects murine from concanavalin A-induced liver injury through inhibition of inflammatory reactions and hepatocyte apoptosis.</a> Biol. Pharm. Bull. 31 (11), 2040-2044 (2008).</li><li>Kaur, G., Tirkey, N., Chopra, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beneficial+effect+of+hesperidin+on+lipopolysaccharide-induced+hepatotoxicity.">Beneficial effect of hesperidin on lipopolysaccharide-induced hepatotoxicity.</a> Toxicology. 226 (2-3), 152-160 (2006).</li><li>Rupertus, K., 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=Major+but+not+minor+hepatectomy+accelerates+engraftment+of+extrahepatic+tumor+cells.">Major but not minor hepatectomy accelerates engraftment of extrahepatic tumor cells.</a> Clin. Exp. Metastasis. 24 (1), 39-48 (2007).</li><li>Selden, C., Casbard, A., Themis, M., Hodgson, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+long-term+survival+of+syngeneic+hepatocytes+in+rat+peritoneum.">Characterization of long-term survival of syngeneic hepatocytes in rat peritoneum.</a> Cell Transplant. 12 (6), 569-578 (2003).</li><li>Tang, T. 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=The+role+of+donor+hepatocytes+and/or+splenocytes+pre-injection+in+reducing+islet+xenotransplantation+rejection.">The role of donor hepatocytes and/or splenocytes pre-injection in reducing islet xenotransplantation rejection.</a> Hepatobiliary. Pancreat. Dis. Int. 2 (3), 344-350 (2003).</li><li>Goto, Y., Ohashi, K., Utoh, R., Yamamoto, M., Okano, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatocyte+transplantation+through+the+hepatic+vein:+a+new+route+of+cell+transplantation+to+the+liver.">Hepatocyte transplantation through the hepatic vein: a new route of cell transplantation to the liver.</a> Cell Transplant. 20 (8), 1259-1270 (2011).</li><li>Gabelein, G., 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=Intrasplenic+or+subperitoneal+hepatocyte+transplantation+to+increase+survival+after+surgically+induced+hepatic+failure?.">Intrasplenic or subperitoneal hepatocyte transplantation to increase survival after surgically induced hepatic failure?.</a> Eur. Surg. Res. 41 (3), 253-259 (2008).</li><li>Bhattacharjee, J., 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=Autologous+NeoHep+Derived+From+Chronic+Hepatitis+B+Virus+Patients'+Blood+Monocytes+by+Upregulation+of+cMET+Signaling.">Autologous NeoHep Derived From Chronic Hepatitis B Virus Patients' Blood Monocytes by Upregulation of cMET Signaling.</a> Stem Cells Transl. Med. , (2016).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Basic surgical skills. Emergency and Essential Surgical Care (EESC) programme. , (2017).</li><li>Pritchett-Corning, K. R., Luo, Y., Mulder, G. B., White, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+rodent+surgery+for+the+new+surgeon.">Principles of rodent surgery for the new surgeon.</a> J. Vis. Exp. (47), (2011).</li><li>Lee, S. M., Schelcher, C., Demmel, M., Hauner, M., Thasler, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+human+hepatocytes+by+a+two-step+collagenase+perfusion+procedure.">Isolation of human hepatocytes by a two-step collagenase perfusion procedure.</a> J. Vis. Exp. (79), (2013).</li><li>Shen, L., Hillebrand, A., Wang, D. Q., Liu, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+primary+culture+of+rat+hepatic+cells.">Isolation and primary culture of rat hepatic cells.</a> J. Vis. Exp. (64), (2012).</li><li>Bhattacharjee, J., 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=Autologous+NeoHep+Derived+from+Chronic+Hepatitis+B+Virus+Patients'+Blood+Monocytes+by+Upregulation+of+c-MET+Signaling.">Autologous NeoHep Derived from Chronic Hepatitis B Virus Patients' Blood Monocytes by Upregulation of c-MET Signaling.</a> Stem Cells Transl. Med. 6 (1), 174-186 (2017).</li><li>Hori, 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=Simple+and+sure+methodology+for+massive+hepatectomy+in+the+mouse.">Simple and sure methodology for massive hepatectomy in the mouse.</a> Ann. Gastroenterol. 24 (4), 307-318 (2011).</li><li>Hori, 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=Simple+and+reproducible+hepatectomy+in+the+mouse+using+the+clip+technique.">Simple and reproducible hepatectomy in the mouse using the clip technique.</a> World J. Gastroenterol. 18 (22), 2767-2774 (2012).</li><li>Vidal, I., Richert, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+nude+mouse+as+model+for+liver+deficiency+study+and+treatment+xenotransplantation.">The nude mouse as model for liver deficiency study and treatment xenotransplantation.</a> Int. J. Hepatol. , 1400147 (2012).</li><li>Mitchell, C., Willenbring, 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+reproducible+and+well-tolerated+method+for+2/3+partial+hepatectomy+in+mice.">A reproducible and well-tolerated method for 2/3 partial hepatectomy in mice.</a> Nat. Protoc. 3 (7), 1167-1170 (2008).</li><li>Ahmed, S. U., 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=Generation+of+subcutaneous+and+intrahepatic+human+hepatocellular+carcinoma+xenografts+in+immunodeficient+mice.">Generation of subcutaneous and intrahepatic human hepatocellular carcinoma xenografts in immunodeficient mice.</a> J. Vis. Exp. (79), e50544 (2013).</li><li>Krikri, 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=Laparoscopic+vs.+open+abdominal+surgery+in+male+pigs:+marked+differences+in+cortisol+and+catecholamine+response+depending+on+the+size+of+surgical+incision.">Laparoscopic vs. open abdominal surgery in male pigs: marked differences in cortisol and catecholamine response depending on the size of surgical incision.</a> Hormones (Athens). 12 (2), 283-291 (2013).</li></ol>

Delvis hepatectomy er en etableret teknik for behandlende lever regenerering, og overdreven hepatectomy er rapporteret at efterligne AHF model. Blandt dyremodeller af Husholdningsfolie af aluminium er gnavere, især mus, de mest undersøgte model. For at opnå en leverskade model i mus, op til en 70% hepatectomy er blevet rapporteret med en god overlevelse sats25,26. Men i nøgen og andre immundefekte mus, en 70% hepatectomy blev rapporteret som fatal og dyr død...

I øjeblikket er hepatocyt transplantation foreslået som et alternativ til hele organtransplantation for behandling af patienter med alvorlige leversygdomme. Det menes, at det kan slå bro over patienter til hele orgel transplantation1. Ud over allogen hepatocytter2, xenogen hepatocytter3 og hepatocytter stammer fra stamceller4 er også ved at blive undersøgt i dyremodeller. I denne sammenhæng, målsøgende og engraftment er potentialet i de transplanterede celler i modtageren et vigtigt kriterium for celle-baserede terapi i akut leversvigt (AHF).
For at undersøge transplantation af hepatocytter eller hepatocyt-lignende celler5, er AHF lavet i en dyremodel af kirurgisk6 eller farmakologiske7 procedurer, efterfulgt af transplantation af celler. At gøre en AHF dyremodel af farmakologiske reagenser, mange hepatotoxins d-galactosamine8, acetaminophen9, tetrachlormethan10, thioacetamide11, Concanavalin A12, LPS13 , osv., har været brugt. Fra denne liste, hver reagens genererer et unikt sæt af funktioner for AHF, men desværre ingen enkelt reagens efterligner de menneskelige AHF. Derudover AHF induceret af hepatotoxins tager lang tid, der sætter dyr under kronisk stress, og reproducerbare resultater er vanskelig at opnå.
På den anden side den kirurgiske procedure af delvis hepatectomy (PHx) er dygtighed afhængige, og reproducerbare resultater er let at opnå efter udvikler nødvendige færdigheder. For at fremkalde AHF ved kirurgisk indgreb alene, er resektion af mere end 70% af leveren påkrævet; dog transplanterede mindre end en 70% hepatectomy kan stadig udnyttes til at studere engraftment og spredning af celler i leveren til at analysere deres terapeutiske kapacitet under leverskader14. Transplantation af hepatocytter har været udført post hepatectomy gennem bughinden15, hale vene16, leverens vene17eller milt18. I øjeblikket, er leverens vene infusion og intrasplenic transplantation af hepatocytter de foretrukne procedurer, så de er nemmere at reproducere.
I dette papir, har vi beskrevet en procedure for en 30% delvis hepatectomy i NOD. SCID (NIK. CB17-Prkdcscid/J) mus, hvor den venstre lap af leveren er skåret ud. Det er efterfulgt af transplantation af 0,2 millioner normal god landbrugspraksis udtrykker mus (C57BL/6-Tg (UBC-NGL) 30Scha/J) hepatocytter samt menneskelig oprindelse NeoHep19 i milten. Denne fremgangsmåde fører til engraftment af de transplanterede celler i leveren. Denne procedure er den mindst invasive og et minimalt smertefulde teknik.

<table border="0" cellpadding="0" cellspacing="0" width="1002">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Gas Anesthesia System</td>
			<td style="width:255px;">Ugo Basile; Italy</td>
			<td style="width:119px;">211000</td>
			<td style="width:217px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Weighing machine</td>
			<td style="width:255px;">Goldtech ; India</td>
			<td style="width:119px;"></td>
			<td>Local Procurement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Biological safety cabinet ( Class I)</td>
			<td style="width:255px;">Kartos international;&#160; India</td>
			<td style="width:119px;"></td>
			<td>Local Procurement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hair Trimmer</td>
			<td>Panasonic ;&#160; Japan&#160;</td>
			<td style="width:119px;">ER-GY10&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Straight operating scissor with sharp /sharp blades</td>
			<td style="width:255px;">Major Surgicals; India</td>
			<td style="width:119px;"></td>
			<td>Local Procurement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps with Serrations</td>
			<td style="width:255px;">Major Surgicals; India</td>
			<td style="width:119px;"></td>
			<td>Local Procurement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro needle holders&#160; straight &#38; curved&#160;</td>
			<td>Mercian ;&#160; England&#160;</td>
			<td style="width:119px;">BS-13-8</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1 ml insulin syringe with 30G *5/16 needles&#160;</td>
			<td>Dispo Van; India</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1 ml syringe with 26 G * 1/2 needle</td>
			<td>BD ; US&#160;</td>
			<td>REF 303060</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nylon Threads&#160;&#160;</td>
			<td style="width:255px;">Mighty ; India</td>
			<td style="width:119px;"></td>
			<td>(1-0) Local Procurement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MERSUTURES 4-0 Sterilised Surgical Needled Suture</td>
			<td style="width:255px;">Ethicon, Johnson &#38; Johnson, India</td>
			<td style="width:119px;">NW 5047</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TRUGUT 76 cm 4-0 absorbable surgical suture</td>
			<td>Sutures India Pvt. Ltd; India</td>
			<td>SN 5048</td>
			<td>Sterilised Surgical Needled Suture Catgut Chromic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Cotton Buds</td>
			<td style="width:255px;">Pure Swabs Pvt Ltd ;&#160; India</td>
			<td style="width:119px;"></td>
			<td>Local Procurement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Surgical Tape</td>
			<td style="width:255px;">3M India ; India</td>
			<td style="width:119px;">1530-1</td>
			<td>Micropore Surgical Tape</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Microtome</td>
			<td style="width:255px;">Histo-Line Laboratories, Italy</td>
			<td style="width:119px;">MRS3500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Shandon Cryotome E Cryostat</td>
			<td style="width:255px;">Thermo Electron Corporation ; US</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Confocal laser scanning microscope</td>
			<td style="width:255px;">Carl Zeiss ; Germany&#160;</td>
			<td style="width:119px;">LSM 510 META</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Bright Field Microscope</td>
			<td style="width:255px;">Olympus, Japan</td>
			<td style="width:119px;">LX51</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:411px;">Automated analyser</td>
			<td>Tulip, Alto Santracruz, India</td>
			<td style="width:119px;">Screen Maaster 3000</td>
			<td>Biochemical analyser for liver functional test</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Flow Cytometer</td>
			<td style="width:255px;">BD ; US&#160;</td>
			<td style="width:119px;">BD FACSverse</td>
			<td>Assesment of presence of cells post transplantation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Veet hair removal cream&#160;</td>
			<td>Reckitt Benckiser , India</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FORANE</td>
			<td style="width:255px;">Abbott ; US</td>
			<td style="width:119px;"></td>
			<td>isoflurane USP 99.9%&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Taxim</td>
			<td style="width:255px;">AlKem ; India</td>
			<td style="width:119px;"></td>
			<td>cefotaxime sodium injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Povidone-Iodine solution&#160;</td>
			<td style="width:255px;">Win-Medicare;&#160; India</td>
			<td style="width:119px;"></td>
			<td>Betadine</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Paraformaldehyde</td>
			<td style="width:255px;">Himedia; India</td>
			<td style="width:119px;">GRM 3660</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:411px;">Iscove&#39;s Modified Dulbecco&#39;s Medium (IMDM)</td>
			<td style="width:255px;">Life technologies, Thermo Fisher scientific ; US</td>
			<td style="width:119px;">12200-036</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Sucrose</td>
			<td style="width:255px;">Sigma ; US</td>
			<td style="width:119px;">S0389</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Tissue-Tek</td>
			<td style="width:255px;">Sakura; US</td>
			<td>25608-930</td>
			<td>O.C.T compound</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">DAPI</td>
			<td style="width:255px;">Himedia; India</td>
			<td style="width:119px;">MB 097</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">anti-Albumin goat Polyclonal</td>
			<td style="width:255px;">Thermo Scientific,Pierce, US</td>
			<td style="width:119px;">PA126081</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">anti-connexin 32/GJB1 Polyclonal</td>
			<td style="width:255px;">abcam, UK</td>
			<td style="width:119px;">ab64609-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">antiGFP rabbit polyclonal&#160;</td>
			<td style="width:255px;">Santa Cruz biotechnology; US</td>
			<td style="width:119px;">SC 8334</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:411px;">Alexa Fluor 594 donkey anti-goat</td>
			<td style="width:255px;">&#160;Molecular Probes , Thermo Fisher Scientific ;&#160; US</td>
			<td style="width:119px;">A11058</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:411px;">Alexa Fluor 488 donkey anti-sheep</td>
			<td style="width:255px;">&#160;Molecular Probes , Thermo Fisher Scientific ;&#160; US</td>
			<td style="width:119px;">A11015</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:411px;">Alexa Fluor 594 chicken anti rabbit</td>
			<td style="width:255px;">&#160;Molecular Probes , Thermo Fisher Scientific ;&#160; US</td>
			<td style="width:119px;">A21442</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:411px;">Goat anti rabbit IgG HRP</td>
			<td style="width:255px;">Invitrogen, Thermo Fisher Scientific; US&#160;</td>
			<td>65-6120</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">anti-Ki67 antibody</td>
			<td style="width:255px;">abcam, UK</td>
			<td style="width:119px;">ab15580</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Antigen Unmasking Solution, Citric acid base</td>
			<td style="width:255px;">Vector laboratories, US</td>
			<td style="width:119px;">H-3300</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">ProLong Diamond antifade mountant</td>
			<td style="width:255px;">Life technologies, Thermo Fisher scientific ; US</td>
			<td style="width:119px;">P36966</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">SGOT (ASAT) KIT</td>
			<td style="width:255px;">Coral Clinical System, India</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">SGPT (ALAT) KIT</td>
			<td style="width:255px;">Coral Clinical System, India</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Alkaline Phosphatase Kit (DEA)</td>
			<td style="width:255px;">Coral Clinical System, India</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Hematoxylin Solution, Mayer&#39;s</td>
			<td style="width:255px;">Sigma ; US</td>
			<td style="width:119px;">MHS16</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Eosin Y solution, alcoholic</td>
			<td style="width:255px;">Sigma ; US</td>
			<td style="width:119px;">HT110132</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">DPX Mountant&#160;</td>
			<td style="width:255px;">Sigma ; US</td>
			<td align="right" style="width:119px;">6522</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">Melonex (Pain Killer)</td>
			<td style="width:255px;">Intas Pharmaceuticals Ltd; India</td>
			<td style="width:119px;"></td>
			<td>Meloxicam injection&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:411px;">DAB enhanced liquid substrate system tetrahydrochloride</td>
			<td style="width:255px;">Sigma ; US</td>
			<td style="width:119px;">D3939</td>
			<td></td>
		</tr>
	</tbody>
</table>

intrasplenic transplantation of hepatocytes after partial hepatectomy in nod.scid mice

Delvis hepatectomy er en alsidig og reproducerbar metode til at studere lever regenerering og effekten af celle-baserede therapeutics i forskellige patologiske tilstande. Delvis hepatectomy letter også øget engraftment og spredning af transplanterede celler af accelererende neovascularization og celle migration til leveren. Her beskriver vi en enkel protokol for udfører 30% hepatectomy og transplantation af celler i milt fra en ikke-overvægtige diabetikere/svær kombineret immundefekte NOD. SCID (NIK. CB17-Prkdcscid/J) mus.
I denne procedure, er to små snit lavet. Den første indsnit er at eksponere og resect venstre lap af leveren, og en anden lille snit er lavet til at afsløre milt for intrasplenic transplantation af celler. Denne procedure kræver ikke nogen specialiserede kirurgiske færdigheder, og det kan være afsluttet i 5-7 minutter med mindre stress og smerter, hurtigere helbredelse og bedre overlevelse. Vi har demonstreret transplantation af hepatocytter isoleret fra en grøn fluorescerende protein (NGL) udtrykker mus (genetisk modificeret C57BL/6-Tg (UBC-NGL) 30Scha/J), samt hepatocyt som celler af human oprindelse (NeoHep) i delvis hepatectomized hilsen. SCID mus.

Hepatocyt spredning efter 30% delvis hepatectomy: Spredning af hepatocytter i de resterende lever efter 30% hepatectomy blev undersøgt ved immunhistokemiske (IHC) farvning til celle spredning markør, Ki-67. Endags-post hepatectomy, mus blev aflivet, de resterende lever kamre var skåret ud og paraffinsnit blev indhentet. Sektionerne var plettet med Ki-67 antistof, efterfulgt af mærkning med peberrod peberrodsperoxidase (HRP)-konjugeret sekundær antistof. Di-Amino benzidin (DAB) blev brugt som substra...

Watch this Scientific Journal Video about Intrasplenic Transplantation of Hepatocytes After Partial Hepatectomy in NOD.SCID Mice at JoVE.com

Intrasplenic Transplantation of Hepatocytes After Partial Hepatectomy in NOD.SCID Mice

Vi har beskrevet en protokol for at udføre delvis hepatectomy (PHx) og celle transplantation via milten i Nik. SCID (NIK. CB17-Prkdcscid/J) mus. I denne protokol, er et snit lavet til at afsløre og resect venstre lap af leveren efterfulgt af et andet snit til intrasplenic transplantation af celler.

Intrasplenic Transplantation af hepatocytter efter delvis Hepatectomy i NOD. SCID mus

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological ...

Research

JoVE Journal

Medicine

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Kirurgiske procedurer for en rottemodel af partiel ortotopisk levertransplantation med hepatisk arteriel Genopbygning

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Heterotopic Cervical Heart Transplantation in Mice

Heterotopisk Livmoderhalskræft hjertetransplantation i mus

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

Visualisering af Vascular og parenkymale Regeneration efter 70% Delvis hepatektomi i normale mus

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Re Arterialized rotte delvis lever Transplantation med en i vivo fartøj-orienterede 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

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

Transplantation af fedtvæv-afledte stamceller ark til at reducere udsivningen efter delvis kolektomi i en rotte Model

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

Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine

Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine

Lentiviral vektor-medieret genterapi af hepatocytter Ex Vivo til autolog Transplantation hos svin

Gene therapy is an ideal choice to cure many inborn errors of metabolism of the liver. Ex-vivo, lentiviral vectors have been used successfully in the ...

Generation of a Rat Model of Acute Liver Failure by Combining 70% Partial Hepatectomy and Acetaminophen

Generation af en rotte model af akut leversvigt ved at kombinere 70% partiel Hepatektomi og acetaminophen

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

Reduceret komplikationer efter arteriel gentilslutning i en rotte model af ortopisk levertransplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Intramyocardial Transplantation of MSC-Loading Injectable Hydrogels after Myocardial Infarction in a Murine Model

Intramyocardial Transplantation af MSC-Loading Injicerbare Hydrogels efter Myocardial Infarkt i en Murine Model

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of ...