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

この作品は、インド政府ニューデリー国立免疫学研究所生物資源科学部から受領したコアによって支えられました。博士バッタチャルジー現住所は消化器科、肝臓と栄養、子供のロサンゼルスの病院です。

このプロトコルで説明されている手順は、ニューデリー国立免疫学研究所の機関の動物倫理委員会によって承認されています。承認のシリアル番号は IAEC #319/13.
注: 一般的な手術手順20と齧歯動物の外科21のための特定のプロトコルに優秀な資源があります。初めて動物の外科手術を行うため、勧め広範囲の練習にダミーに手術動...

<ol>
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肝部分切除は調査の肝再生に確立された技術と過剰な切除は AHF モデルを模倣に報告されました。AHF の動物モデルは、齧歯動物、特にマウスは最も研究のモデルがあります。70% 肝切除までのマウス肝障害モデルを取得するには、良い生存率25,26で報告されています。しかし裸や他の免疫不全マウスは、70% 肝切除を致命的なものとして報告された、2...

現在、重度の肝障害を有する患者の治療のための全体の臓器移植に代わるものとして肝細胞移植を提案する.それが全臓器移植1患者を埋めることができるといわれています。同種肝2、に加えてまた異種肝3と4幹細胞由来肝細胞を動物モデルで検討されています。このコンテキスト、ホーミングと生着受信者に移植細胞の潜在性は急性肝不全 (AHF) で基づく細胞療法の重要な基準です。
肝細胞の肝細胞様細胞5移植を調査、AHF が手術6か細胞移植薬理7手続、動物モデルで作成されます。薬理学的試薬、コンカナバリン A12、リポ多糖13, チオアセトアミド11四塩化炭素10アセトアミノフェン9d-ガラクトサミン8など多くの hepatotoxins によって AHF 動物モデルを作成するには、等.、使用されています。このリストからすべての試薬、AHF のユニークな一連の機能を生成しますが、単一試薬を人間 AHF 模倣ない残念なことに。また、hepatotoxins による AHF 慢性的なストレス下での動物を置く、長い時間がかかる、再現性のある結果が得にくい。
その一方で、肝部分切除 (PHx) の手術はスキル依存と再現性のある結果が必要なスキルを開発したら、入手しやすい。単独で外科的介入によって AHF を誘導して、肝臓の 70% 以上の切除が必要です。ただし、70% 肝切除は生着、増殖研究にまだ利用することができます未満は肝臓障害14中治療能力の分析の肝細胞を移植しました。肝細胞の移植は、腹膜15、16尾静脈、肝静脈17、または脾臓18を介して実行されるポスト肝切除をされています。現在、肝静脈注入と脾内移植肝細胞の最寄りの手順では、簡単に再現しています。
本稿でうなずきで 30% 部分肝切除のための手順を説明しました。SCID (うなずく。CB17-Prkdcscid/J) マウス肝臓の左葉が切除されます。それは、人間の起源は、脾臓で NeoHep19と同様、20 万の GFP 発現マウス (C57BL/6-Tg (UBC GFP) 30Scha/J) 肝細胞の移植が続きます。この手順は、肝臓移植細胞の生着につながります。この手順は、少なくとも侵襲的な少なくとも痛みを伴う方法です。

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			<td>REF 303060</td>
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			<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

肝切除は、肝再生と各種病態における細胞による治療の効果を研究する汎用性と再現性のある方法です。肝部分切除は加速新生血管と肝臓への細胞移行によってまた増加移植と移植細胞の増殖を促進します。ここでは、免疫不全のうなずきを組み合わせた非肥満糖尿病/重度の脾臓で 30% 肝切除や細胞移植を実行するための単純なプロトコルについて述べる.SCID (うなずく。CB17-Prkdcscid/J) マウス。
この手順では、2 つの小さな切開が作られています。最初の切開を公開し, 肝左葉を切除、脾内移植細胞の脾臓を公開するもう一つの小さな切開を行った。この手順は、すべての専門的な手術のスキルを必要はありません、少ないストレスと痛み、高速回復、生存率と 5-7 分で完了することができます。マウス (トランスジェニック C57BL/6-Tg (UBC GFP) 30Scha/J)、として部分的肝切除会釈で人間の起源 (NeoHep) の細胞のような肝細胞を表現する緑色蛍光タンパク質 (GFP) から分離された肝細胞の移植を行った。SCID マウス。

30% 肝部分切除後の肝細胞増殖:細胞増殖マーカー、Ki 67 の染色 (IHC) を免疫組織化学によって 30% 肝切除後肝残りの肝細胞の増殖を調べた。1 日ポスト肝マウスを安楽死させ、残りの肝葉摘出、およびパラフィン切片が得られました。西洋わさびペルオキシダーゼ (HRP) 共役二次抗体とラベリングに続いて、Ki 67 抗体染色しました。・ ディ ・ アミノ ベンジジン (軽打) は、染色の細?...

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

ノドの部分肝切除 (PHx) と脾臓を介して細胞移植を実行するためのプロトコルを説明しました。SCID (うなずく。CB17-Prkdcscid/J) マウス。このプロトコルを公開し、脾細胞移植のための別の切開に続いて肝臓の左葉を切除する切開します。

うなずくにおける肝切除後肝脾内移植。SCID マウス

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

Research

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Medicine

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

肝動脈再建を用いた部分同所性肝移植のラットモデルのための外科的処置

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

マウスにおける異子宮頸心臓移植

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

正常マウスで70％肝切除後の血管と実質再生の可視化

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

再門脈部分肝移植のラット体内血管指向 70% 肝切除

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

ラットモデルにおける部分的な術後後の漏れを減らすために脂肪組織由来幹細胞シートの移植

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

豚に自家移植用肝細胞Ex Vivoのレンチ ウイルス媒介性遺伝子治療

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

70%部分肝切除術とアセトアミノフェンを組み合わせた急性肝不全ラットモデルの生成

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

ラット型異形肝移植モデルにおける動脈リコネクション後の合併症の減少

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

マウスモデルにおける心筋梗塞後のMSC負荷注入性ヒドロゲルの筋内移植

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