National Center of Excellence in Molecular Biology, 87-West Canal Bank Road, University of the Punjab, Lahore, Pakistan.
Differentiation. 2011 Jan;81(1):42-8. doi: 10.1016/j.diff.2010.08.005. Epub 2010 Oct 12.
Liver failure represents a serious challenge for cell based therapies. Mesenchymal stem cells (MSCs) possess potential for regeneration of fibrotic liver; however, there is a dire need to improve their hepatic differentiation. This study examines a pretreatment strategy to augment the differentiation potential of MSCs towards hepatic lineage. MSCs were isolated from C57BL/6 wild type mice and were characterized by flow cytometry for CD44 (92.4%), CD90 (96.6%), CD105 (94.7%), CD45 (0.8%) and CD34 (1.4%) markers. To improve the differentiation potential of MSCs towards hepatic lineage, cells were pretreated with injured liver tissue in an in-vitro model, which resulted in high expression of albumin, cytokeratin 8, 18, TAT and HNF1α as compared to untreated MSCs. The efficacy of pretreated MSCs was evaluated by preparing in-vivo mouse model with liver fibrosis by intraperitoneal administration of CCl(4). Pretreated MSCs were transplanted in the left lateral lobe of mice with liver fibrosis and showed enhanced localization and differentiation abilities after 1 month. The expression for cytokeratin 8, 18, albumin and Bcl-xl was up-regulated and that of HGF, Bax and Caspase- 3 was down-regulated in animals transplanted with pretreated MSCs. Sirus red staining also confirmed a significant reduction in the fibrotic area in liver tissue transplanted with pretreated MSCs as compared to untreated MSCs and was concomitant with improved serum levels of bilirubin and alkaline phosphatase (ALP). Therefore, it was concluded that pretreatment with injured liver tissue augment homing and hepatic differentiation abilities of MSCs and provides an improved procedure for the treatment of liver fibrosis.
肝衰竭是细胞治疗面临的严重挑战。间充质干细胞（MSCs）具有纤维化肝脏再生的潜力；然而，急需提高其向肝系分化的能力。本研究探讨了一种预处理策略，以增强 MSCs 向肝系分化的潜能。MSCs 从小鼠的 C57BL/6 野生型中分离出来，并通过流式细胞术对 CD44（92.4%）、CD90（96.6%）、CD105（94.7%）、CD45（0.8%）和 CD34（1.4%）标志物进行了鉴定。为了提高 MSCs 向肝系分化的潜能，将细胞在体外模型中用受损的肝组织进行预处理，与未经处理的 MSCs 相比，白蛋白、细胞角蛋白 8、18、TAT 和 HNF1α 的表达明显升高。通过腹腔内给予 CCl(4)制备肝纤维化的体内小鼠模型来评估预处理 MSCs 的功效。将预处理的 MSCs 移植到肝纤维化的小鼠左侧外侧叶中，1 个月后显示出增强的定位和分化能力。在移植预处理 MSCs 的动物中，细胞角蛋白 8、18、白蛋白和 Bcl-xl 的表达上调，HGF、Bax 和 Caspase-3 的表达下调。Sirus red 染色也证实，与未处理的 MSCs 相比，预处理 MSCs 移植的肝组织中纤维化面积显著减少，同时血清胆红素和碱性磷酸酶（ALP）水平也得到改善。因此，结论是用受损的肝组织预处理可以增强 MSCs 的归巢和肝分化能力，并为肝纤维化的治疗提供了一种改进的方法。
