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部分肝切除术后NOD.SCID小鼠的脾内肝细胞移植

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

作者信息

Das Barun, Bhattacharjee Jashdeep, Mishra Alaknanda, Jain Kshama, Iyer Srikanth, Kesarwani Ashwani, Sahu Parul, Sinha Prakriti, Nagarajan Perumal, Upadhyay Pramod

机构信息

National Institute of Immunology.

National Institute of Immunology;

出版信息

J Vis Exp. 2018 Feb 10(132):56018. doi: 10.3791/56018.

Abstract

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological conditions. Partial hepatectomy also facilitates the increased engraftment and proliferation of transplanted cells by accelerating neovascularization and cell migration towards the liver. Here, we describe a simple protocol for performing 30% hepatectomy and transplantation of cells in the spleen of a non-obese diabetic/severe combined immunodeficient NOD.SCID (NOD.CB17-Prkdc/J) mouse. In this procedure, two small incisions are made. The first incision is to expose and resect the left lobe of the liver, and another small incision is made to expose the spleen for the intrasplenic transplantation of cells. This procedure does not require any specialized surgical skills, and it can be completed in 5-7 minutes with less stress and pain, faster recovery, and better survival. We have demonstrated the transplantation of hepatocytes isolated from a green fluorescent protein (GFP) expressing mouse (Transgenic C57BL/6-Tg (UBC-GFP) 30Scha/J), as well as hepatocyte like cells of human origin (NeoHep) in partially hepatectomized NOD.SCID mice.

摘要

部分肝切除术是一种通用且可重复的方法,用于研究肝再生以及基于细胞的治疗方法在各种病理条件下的效果。部分肝切除术还通过加速新血管形成和细胞向肝脏的迁移,促进移植细胞的植入和增殖。在此,我们描述一种在非肥胖糖尿病/重症联合免疫缺陷NOD.SCID(NOD.CB17-Prkdc/J)小鼠脾脏中进行30%肝切除术和细胞移植的简单方案。在该手术中,需做两个小切口。第一个切口用于暴露并切除肝脏左叶,另一个小切口用于暴露脾脏以进行脾内细胞移植。此手术不需要任何特殊的外科手术技巧,可在5至7分钟内完成,应激和疼痛较小,恢复更快,存活率更高。我们已在部分肝切除的NOD.SCID小鼠中证明了从表达绿色荧光蛋白(GFP)的小鼠(转基因C57BL/6-Tg (UBC-GFP) 30Scha/J)分离的肝细胞以及人源肝样细胞(NeoHep)的移植。

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