Department of Surgery, Campus Charité Mitte and Campus Virchow-Klinikum, Charité - Universitaetsmedizin, Berlin, Germany.
Department of Surgery, Campus Charité Mitte and Campus Virchow-Klinikum, Charité - Universitaetsmedizin, Berlin, Germany.
J Surg Res. 2019 Jul;239:191-200. doi: 10.1016/j.jss.2019.02.010. Epub 2019 Mar 4.
Mesenchymal stem cells (MSCs) have been suggested to augment liver regeneration after surgically and pharmacologically induced liver failure. To further investigate this we processed human bone marrow-derived MSC according to good manufacturing practice (GMP) and tested those cells for their modulatory capacities of metabolic alterations and liver regeneration after partial hepatectomy in BALB/c nude mice.
Human MSCs were obtained by bone marrow aspiration of healthy donors as in a previously described GMP process. Transgenic GFP-MSCs were administered i.p. 24 h after 70% hepatectomy in BALB/c nude mice, whereas control mice received phosphate-buffered saline. Mice were sacrificed 2, 3, and 5 d after partial hepatectomy. Blood and organs were harvested and metabolic alterations as well as liver regeneration subsequently assessed by liver function tests, multianalyte profiling immunoassays, histology, and immunostaining.
Hepatocyte and sinusoidal endothelial cell proliferation were significantly increased after partial hepatectomy in mice receiving MSC compared to control mice (Hepatocyte postoperative day 3, P < 0.01; endothelial cell postoperative day 5, P < 0.05). Hepatocyte fat accumulation correlated inversely with hepatocyte proliferation (r = 0.4064, P < 0.01) 2 d after partial hepatectomy, with mice receiving MSC being protected from severe fat accumulation. No GFP-positive cells could be detected in the samples. Serum levels of IL-6, HGF, and IL-10 were significantly decreased at day 3 in mice receiving MSC when compared to control mice (P < 0.05). Relative body weight loss was significantly attenuated after partial hepatectomy in mice receiving MSC (2 d and 3 d, both P < 0.001) with a trend toward a faster relative restoration of liver weight, when compared to control mice.
Human bone marrow-derived MSC attenuate metabolic alterations and improve liver regeneration after partial hepatectomy in BALB/c nude mice. Obtained results using GMP-processed human MSC suggest functional links between fat accumulation and hepatocyte proliferation, without any evidence for cellular homing. This study using GMP-proceeded MSC has important regulatory implications for an urgently needed translation into a clinical trial.
间充质干细胞（MSCs）被认为可以增强手术和药物诱导的肝衰竭后的肝再生。为了进一步研究这一点，我们根据良好生产规范（GMP）处理了人骨髓来源的 MSC，并在 BALB/c 裸鼠的部分肝切除后测试了这些细胞对代谢改变和肝再生的调节能力。
人 MSC 通过健康供体的骨髓抽吸获得，如先前描述的 GMP 过程。在 BALB/c 裸鼠 70%肝切除后 24 小时，经腹腔注射转基因 GFP-MSC，而对照小鼠给予磷酸盐缓冲盐水。部分肝切除后 2、3 和 5 天处死小鼠。采集血液和器官，通过肝功能试验、多分析物谱免疫分析、组织学和免疫染色评估代谢改变和肝再生。
与对照组相比，接受 MSC 的小鼠部分肝切除后肝细胞和窦内皮细胞增殖明显增加（肝细胞术后第 3 天，P<0.01；内皮细胞术后第 5 天，P<0.05）。肝细胞脂肪堆积与肝细胞增殖呈负相关（r=0.4064，P<0.01），部分肝切除后 2 天，接受 MSC 的小鼠受到严重脂肪堆积的保护。在样本中未检测到 GFP 阳性细胞。与对照组相比，接受 MSC 的小鼠术后第 3 天血清中 IL-6、HGF 和 IL-10 水平显著降低（P<0.05）。接受 MSC 的小鼠部分肝切除后，相对体重减轻明显减轻（2 天和 3 天，均 P<0.001），与对照组相比，肝重相对恢复的趋势更快。
人骨髓来源的 MSC 可减轻 BALB/c 裸鼠部分肝切除后的代谢改变并改善肝再生。使用 GMP 处理的人 MSC 获得的结果表明，脂肪堆积和肝细胞增殖之间存在功能联系，而没有任何细胞归巢的证据。这项使用 GMP 处理的 MSC 的研究对迫切需要转化为临床试验具有重要的监管意义。
