急性肝衰竭模型猪微环境对脂肪间充质干细胞生物学特性的影响  

Microenvironment of acute liver failure pig effects on the biological characteristics of adipose-derived mesenchymal stem cells

作　　者：郭瑞红 潘小平 侯晓丽 张悦 江夏薇 汪恬甜 刘寿荣 GUO Ruihong;PAN Xiaoping;HOU Xiaoli;ZHANG Yue;JIANG Xiawei;WANG Tian-tian;LIU Shourong(Zhejiang University of Traditional Chinese Medicine, Hangzhou, Zhejiang province, 310053, China;College of Basic Medical Sciences, Zhejiang University of Traditional Chinese Medicine, Hangzhou, Zhejiang province, 310053, China;Academy of Traditional Chinese Medicine, Zhejiang University of Traditional Chinese Medicine, Hangzhou, Zhejiang province, 310053, China;Department of Hepatology, Hangzhou Xixi Hospital Affiliated to Zhejiang University of Traditional Chinese Medicine, Hangzhou, Zhejiang province, 310014, China)

机构地区：[1]浙江中医药大学,杭州310053 [2]浙江中医药大学基础医学院,杭州310053 [3]浙江中医药大学中医药科学院,杭州310053 [4]浙江中医药大学附属杭州西溪医院肝病科,杭州310014

出　　处：《浙江中西医结合杂志》2019年第2期100-105,175,共7页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的探究急性肝衰竭(ALF)猪脂肪来源的间充质干细胞(ADMSCs)的生物学活性,为ALF-ADMSCs是否具有潜在的自体移植潜能提供实验依据。方法分别获取急性肝衰竭猪来源的ADMSCs和正常猪来源的ADMSCs (Nor-ADMSCs),进行原代分离和培养。观察第1代与第5代ALF-ADMSCs的细胞形态,CCK8法检测ALF-ADMSCs增殖能力,检测和分析其诱导分化为成骨细胞、脂肪细胞、成软骨细胞、成肝细胞的能力,以及分析ALF-ADMSCs和Nor-ADMSCs中肝脏特异性基因的表达。结果成功分离培养ALF-ADMSCs和Nor-ADMSCs,第1代与第5代ALFADMSCs细胞形态均与Nor-ADMSCs相似。ALF-ADMSCs和Nor-ADMSCs均高表达间充质干细胞表面标记CD105和CD90,不表达或低表达造血干细胞表面标记CD14。ALF-ADMSCs与NorADMSCs增殖能力差异无统计学意义(P>0.05)。与Nor-ADMSCs相比较,ALF-ADMSCs诱导分化为成骨细胞、脂肪细胞、软骨细胞、肝样细胞的能力差异无统计学意义(P>0.05)。肝细胞生长因子(HGF)在ALF-ADMSCs中表达量比Nor-ADMSCs高(12.48±0.59)倍。结论 ALF-ADMSCs具有正常的干细胞特征与生物学特性,没有因ALF微环境而改变,且其高表达肝细胞生长因子,或可作为干细胞移植治疗终末期肝病的理想细胞源。Objective To explore the biological characteristics of adipose-derived mesenchymal stem cells collected from pigs with acute liver failure(ALF-ADMSCs), in order to providing an experimental basis for the potential application of ADMSCs autologous transplantation in patients with end-stage liver disease. Methods ADMSCs isolated from acute liver failure pig and from a normal pig(Nor-ADMSCs) were primary cultured, respectively. The morphology of the first and fifth generation ALF-ADMSCs was observed. The cell proliferation of ALF-ADMSCs was detected by CCK8. Also, its ability to induce and differentiate into osteoblasts, adipocytes, chondrocytes and hepatoblasts were evaluated. The expression of liver-specific genes in ALF-ADMSCs and Nor-ADMSCs were analyzed. Results ALF-ADMSCs and Nor-ADMSCs were successfully isolated and cultured. The morphology of the first and fifth generation ALF-ADMSCs was similar to that of Nor-ADMSCs. Both ALF-ADMSCs and Nor-ADMSCs highly expressed the surface marker CD105 and CD90 of mesenchymal stem cells, but did not express or lowly expressed the surface marker CD14 of hematopoietic stem cells. There was no difference in proliferation ability between ALF-ADMSCs and Nor-ADMSCs. Compared to Nor-ADMSCs, ALF-ADMSCs have comparable ability to induce differentiation into osteoblasts, adipocytes, chondrocytes and hepatocyte-like cells. The expression levels of hepatocyte growth factor(HGF) in ALF-ADMSCs was 12.48±0.59 times higher than that of Nor-ADMSCs. Conclusion The results suggested that the stem cell features and biological characteristics of ALF-ADMSCs could remain under the acute liver failure microenvironment. ALF-ADMSCs highly expressed HGF, which indicated that it would be an ideal stem cell source for autologous transplantation in treating end-stage liver disease.

关 键 词：巴马香猪 肝衰竭 脂肪间充质干细胞 分化潜能 生物学特性 自体移植 

分 类 号：R575.3[医药卫生—消化系统] R-332[医药卫生—内科学]