猪的脂肪选择：肝源性分化间充质干细胞用于转化研究。

A fat option for the pig: hepatocytic differentiated mesenchymal stem cells for translational research.

机构信息

University Hospital Leipzig, Department of Visceral, Transplantation, Thoracic and Vascular Surgery, Liebigstraße 21, Leipzig D-04103, Germany.

University Hospital Leipzig, Department of Visceral, Transplantation, Thoracic and Vascular Surgery, Liebigstraße 21, Leipzig D-04103, Germany; TRM, Translational Centre for Regenerative Medicine, Philipp-Rosenthal-Str. 55, Leipzig D-04103, Germany.

出版信息

Exp Cell Res. 2014 Feb 15;321(2):267-75. doi: 10.1016/j.yexcr.2013.10.018. Epub 2013 Nov 4.

DOI:10.1016/j.yexcr.2013.10.018

PMID:24200501

Abstract

STUDY BACKGROUND

Extended liver resection is the only curative treatment option of liver cancer. Yet, the residual liver may not accomplish the high metabolic and regenerative capacity needed, which frequently leads to acute liver failure. Because of their anti-inflammatory and -apoptotic as well as pro-proliferative features, mesenchymal stem cells differentiated into hepatocyte-like cells might provide functional and regenerative compensation. Clinical translation of basic research requires pre-clinical approval in large animals. Therefore, we characterized porcine mesenchymal stem cells (MSC) from adipose tissue and bone marrow and their hepatocyte differentiation potential for future assessment of functional liver support after surgical intervention in the pig model.

METHODS

Mesenchymal surface antigens and multi-lineage differentiation potential of porcine MSC isolated by collagenase digestion either from bone marrow or adipose tissue (subcutaneous/visceral) were assessed by flow cytometry. Morphology and functional properties (urea-, glycogen synthesis and cytochrome P450 activity) were determined during culture under differentiation conditions and compared with primary porcine hepatocytes.

RESULTS

MSC from porcine adipose tissue and from bone marrow express the typical mesenchymal markers CD44, CD29, CD90 and CD105 but not haematopoietic markers. MSC from both sources displayed differentiation into the osteogenic as well as adipogenic lineage. After hepatocyte differentiation, expression of CD105 decreased significantly and cells adopted the typical polygonal morphology of hepatocytes. Glycogen storage was comparable in adipose tissue- and bone marrow-derived cells. Urea synthesis was about 35% lower in visceral than in subcutaneous adipose tissue-derived MSC. Cytochrome P450 activity increased significantly during differentiation and was twice as high in hepatocyte-like cells generated from bone marrow as from adipose tissue.

CONCLUSION

The hepatocyte differentiation of porcine adipose tissue-derived MSC was shown for the first time yielding hepatocyte-like cells with specific functions similar in bone marrow and subcutaneous adipose tissue-derived MSC. That makes them good pre-clinical candidates for supportive approaches after liver resection in the pig.

摘要

研究背景

扩大肝切除术是肝癌唯一的治愈性治疗选择。然而，残余的肝脏可能无法完成所需的高代谢和再生能力，这常常导致急性肝功能衰竭。间充质干细胞分化为肝细胞样细胞具有抗炎、抗凋亡和促增殖的特性，可能提供功能和再生补偿。基础研究的临床转化需要在大型动物中进行临床前批准。因此，我们对来自脂肪组织和骨髓的猪间充质干细胞（MSC）及其向肝细胞分化的潜能进行了表征，以便在猪模型中评估手术干预后对功能性肝支持的作用。

方法

通过胶原酶消化从骨髓或脂肪组织（皮下/内脏）中分离的猪 MSC，通过流式细胞术评估间充质表面抗原和多能分化潜能。在分化条件下培养期间，确定形态和功能特性（尿素、糖原合成和细胞色素 P450 活性），并与原代猪肝细胞进行比较。

结果

来自猪脂肪组织和骨髓的 MSC 表达典型的间充质标志物 CD44、CD29、CD90 和 CD105，但不表达造血标志物。两种来源的 MSC 均能分化为成骨细胞和脂肪细胞谱系。在向肝细胞分化后，CD105 的表达显著降低，细胞呈现出典型的肝细胞多角形形态。糖原储存量在脂肪组织衍生和骨髓衍生细胞中相当。内脏脂肪组织衍生的 MSC 的尿素合成比皮下脂肪组织衍生的 MSC 低约 35%。细胞色素 P450 活性在分化过程中显著增加，并且骨髓衍生的肝细胞样细胞的活性是脂肪组织衍生的细胞的两倍。