Moriya Kei, Yoshikawa Masahide, Saito Ko, Ouji Yukiteru, Nishiofuku Mariko, Hayashi Noriko, Ishizaka Shigeaki, Fukui Hiroshi
Division of Hepatology, Third Department of Internal Medicine, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan.
World J Gastroenterol. 2007 Feb 14;13(6):866-73. doi: 10.3748/wjg.v13.i6.866.
To transplant undifferentiated embryonic stem (ES) cells into the spleens of carbon tetrachloride (CCl4)-treated mice to determine their ability to differentiate into hepatocytes in the liver.
CCl4, 0.5 mL/kg body weight, was injected into the peritoneum of C57BL/6 mice twice a week for 5 wk. In group 1 (n=12), 1 x 10(5) undifferentiated ES cells (0.1 mL of 1 x 10(6)/mL solution), genetically labeled with GFP, were transplanted into the spleens 1 d after the second injection. Group 2 mice (n=12) were injected with 0.2 mL of saline twice a week, instead of CCl4, and the same amount of ES cells was transplanted into the spleens. Group 3 mice (n=6) were treated with CCl4 and injected with 0.1 mL of saline into the spleen, instead of ES cells. Histochemical analyses of the livers were performed on post-transplantation d (PD) 10, 20, and 30.
Considerable numbers of GFP-immunopositive cells were found in the periportal regions in group 1 mice (CCl4-treated) on PD 10, however, not in those untreated with CCl4 (group 2). The GFP-positive cells were also immunopositive for albumin (ALB), alpha-1 antitrypsin, cytokeratin 18, and hepatocyte nuclear factor 4 alpha on PD 20. Interestingly, most of the GFP-positive cells were immunopositive for DLK, a hepatoblast marker, on PD 10. Although very few ES-derived cells were demonstrated immunohistologically in the livers of group 1 mice on PD 30, improvements in liver fibrosis were observed. Unexpectedly, liver tumor formation was not observed in any of the mice that received ES cell transplantation during the experimental period.
Undifferentiated ES cells developed into hepatocyte-like cells with appropriate integration into tissue, without uncontrolled cell growth.
将未分化的胚胎干细胞（ES细胞）移植到四氯化碳（CCl4）处理的小鼠脾脏中，以确定其在肝脏中分化为肝细胞的能力。
每周两次向C57BL/6小鼠腹腔注射0.5 mL/kg体重的CCl4，持续5周。在第1组（n = 12）中，在第二次注射后1天，将1×10(5)个用绿色荧光蛋白（GFP）进行基因标记的未分化ES细胞（0.1 mL的1×10(6)/mL溶液）移植到脾脏中。第2组小鼠（n = 12）每周两次注射0.2 mL生理盐水，而非CCl4，并将相同数量的ES细胞移植到脾脏中。第3组小鼠（n = 6）用CCl4处理，并向脾脏注射0.1 mL生理盐水，而非ES细胞。在移植后第10、20和30天对肝脏进行组织化学分析。
在移植后第10天，第1组（CCl4处理）小鼠的门静脉周围区域发现了大量GFP免疫阳性细胞，而未用CCl4处理的小鼠（第2组）中未发现。在移植后第20天，GFP阳性细胞对白蛋白（ALB）、α1抗胰蛋白酶、细胞角蛋白18和肝细胞核因子4α也呈免疫阳性。有趣的是，在移植后第10天，大多数GFP阳性细胞对成肝细胞标志物DLK呈免疫阳性。尽管在移植后第30天，第1组小鼠肝脏中通过免疫组织化学检测到的ES来源细胞很少，但观察到肝纤维化有所改善。出乎意料的是，在实验期间接受ES细胞移植的任何小鼠中均未观察到肝肿瘤形成。
未分化的ES细胞发育为类似肝细胞的细胞，并适当整合到组织中，没有不受控制的细胞生长。
