Use of Decellularized Porcine Liver for Engineering Humanized Liver Organ
Use of Decellularized Porcine Liver for Engineering Humanized Liver Organ
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DOI：
10.1016/j.jss.2011.09.033
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发表时间：
2012-03-01
影响因子：
2.2
通讯作者：
Gauthier, Polly K.
中科院分区：
文献类型：
--
作者：
Barakat, Omar;Abbasi, Shahrzad;Gauthier, Polly K.
背景资料。需要新的生物人工肝设备来补充为终末期肝病患者提供的有限的器官捐赠者。在此，我们报道了一项旨在通过将与胎儿星状细胞(HFSC)共培养的中期人胎肝细胞(HFH)移植到猪肝脱细胞基质中来开发人源化猪肝的初步研究结果。取19头约克夏猪的缺血肝。用阴离子洗涤剂(十二烷基硫酸钠)去除肝脏细胞。将3种脱细胞猪肝基质分别接种3.5×10(8)和1×10(9)的HFSC和HFH，分别灌流3、7和13d，观察移植细胞在灌流过程中和灌流后的代谢和合成活性。脱细胞基质的免疫组织化学检查显示，核物质被去除，结构完整，细胞外基质(ECM)蛋白保留。在再细胞基质的灌流过程中，代谢参数的测量(即氧浓度、葡萄糖消耗、乳酸和尿素的产生)表明代谢活跃。人血清白蛋白平均浓度为29.48+/-7.4微克/毫升。免疫组织化学分析显示细胞分化为成熟肝细胞。此外，40%的移植细胞处于活跃的增殖状态，只有不到30%的细胞发生凋亡。我们发现，我们的脱细胞方案成功地去除了猪肝的细胞成分，同时保留了天然的结构和大多数细胞外基质蛋白。我们还证明了脱细胞基质在持续灌流系统中支持和诱导植入的HfH的表型成熟的能力。(C)2012 Elsevier Inc.保留所有权利。
Background. New bioartificial liver devices are needed to supplement the limited supply of organ donors available for patients with end-stage liver disease. Here, we report the results of a pilot study aimed at developing a humanized porcine liver by transplanting second trimester human fetal hepatocytes (Hfh) co-cultured with fetal stellate cells (Hfsc) into the decellularized matrix of a porcine liver.Material and Methods. Ischemic livers were removed from 19 Yorkshire swine. Liver decellularization was achieved by an anionic detergent (SDS). The decellularized matrix of three separate porcine liver matrices was seeded with 3.5 x 10(8) and 1 x 10(9) of Hfsc and Hfh, respectively, and perfused for 3, 7, and 13 d. The metabolic and synthetic activities of the engrafted cells were assessed during and after perfusion.Results. Immunohistologic examination of the decellularized matrix showed removal of nuclear materials with intact architecture and preserved extracellular matrix (ECM) proteins. During perfusion of the recellularized matrices, measurement of metabolic parameters (i.e., oxygen concentration, glucose consumption, and lactate and urea production) indicated active metabolism. The average human albumin concentration was 29.48 +/- 7.4 mu g/mL. Immunohistochemical analysis revealed cell differentiation into mature hepatocytes. Moreover, 40% of the engrafted cells were actively proliferating, and less than 30% of cells were apoptotic.Conclusion. We showed that our decellularization protocol successfully removed the cellular components of porcine livers while preserving the native architecture and most ECM protein. We also demonstrated the ability of the decellularized matrix to support and induce phenotypic maturation of engrafted Hfh in a continuously perfused system. (C) 2012 Elsevier Inc. All rights reserved.