Brückner S, Tautenhahn H-M, Winkler S, Stock P, Jonas S, Dollinger M, Christ B
University Hospital Leipzig, Department of Visceral, Transplantation, Thoracic and Vascular Surgery, Leipzig, Germany.
Transplant Proc. 2013 Jun;45(5):2056-8. doi: 10.1016/j.transproceed.2013.01.101.
Mesenchymal stem cells (MSC) isolated from bone marrow and differentiated into hepatocyte-like cells have increasingly gained attention for clinical cell therapy of liver diseases because of their high regenerative capacity. They are available from bone marrow aspirates of the os coxae after puncture of the crista iliaca or from bone marrow "surgical waste" gained from amputations or knee and hip operations. Thus, the aim of the study was to demonstrate whether these pBM-MSC (porcine bone marrow-derived mesenchymal stem cells) displayed mesenchymal features and hepatocyte differentiation potential. MSC were isolated either from crista iliaca punctures or after sampling and collagenase digestion of bone marrow from the os femoris. Mesenchymal features were assessed by flow cytometry for specific surface antigens and their ability to differentiate into at least 3 lineages. Functional properties, such as urea or glycogen synthesis and cytochrome P450 activity, as well as the cell morphology were examined during hepatocyte differentiation. pBM-MSC from both sources lacked the hematopoietic markers CD14 and CD45 but expressed the typical mesenchymal markers CD44, CD29, CD90, and CD105. Both cell types could differentiate into adipocyte, osteocyte, and hepatocyte lineages. After hepatocyte differentiation, CD105 expression decreased significantly and cells changed morphology from fibroblastoid into polygonal, displaying significantly increased glycogen storage, urea synthesis, and cytochrome activity. pBM-MSC from various sources were identical in respect to their mesenchymal features and their hepatocyte differentiation potential. Hence, long bones might be a particularly useful resource to isolate bone marrow mesenchymal stem cells for transplantation.
从骨髓中分离并分化为肝细胞样细胞的间充质干细胞（MSC），因其具有较高的再生能力，在肝脏疾病的临床细胞治疗中越来越受到关注。它们可从髂嵴穿刺后的髋骨骨髓抽吸物中获取，也可从截肢手术或膝关节及髋关节手术中获得的骨髓“手术废弃物”中获取。因此，本研究的目的是证明这些猪骨髓来源的间充质干细胞（pBM-MSC）是否具有间充质特征和肝细胞分化潜能。MSC要么从髂嵴穿刺中分离，要么从股骨骨髓取样并经胶原酶消化后分离。通过流式细胞术评估间充质特征，检测特定表面抗原以及它们分化为至少3种谱系的能力。在肝细胞分化过程中，检测其功能特性，如尿素或糖原合成以及细胞色素P450活性，同时观察细胞形态。来自两种来源的pBM-MSC均缺乏造血标志物CD14和CD45，但表达典型的间充质标志物CD44、CD29、CD90和CD105。两种细胞类型均可分化为脂肪细胞、骨细胞和肝细胞谱系。肝细胞分化后，CD105表达显著降低，细胞形态从成纤维样变为多边形，糖原储存、尿素合成和细胞色素活性显著增加。不同来源的pBM-MSC在间充质特征和肝细胞分化潜能方面是相同的。因此，长骨可能是分离用于移植的骨髓间充质干细胞的特别有用的资源。
