摘要
瞄准:学习人的脐带血干细胞(HUCBSC ) 的条件和潜力区分进 hepatocytes 在活体内或在试管内。方法:在人的脐带血的房间文化研究干细胞(HUCBSC ) 区别,人的脐带血单音的原子房间(HUCBMNC ) 被密度坡度远心沉淀分开。成纤维细胞生长因素(FGF ) 和 hepatocyte 生长因素(HGF ) 和胎儿的肝的上层清液在劝诱的组被增加。仅仅 FGF 在控制组被增加。在每个组的 HUCBMNC 的扩大和区别被观察。人的高山哈 fetoprotein (法新社) 和白朊(白长袍的) 被免疫组织化学检测。在动物实验,有在四氯化碳(CCL4 ) 以后的尖锐肝的损害的幸存 SD 老鼠注射 48 h 随机被划分成三个组。在组 A 的老鼠与人的脐带血浆液被对待。在组 B 的老鼠与 HUCBMNC 移植被对待。在组 C 的老鼠与移植为 7 d 由腹膜内出租机动三轮车磷酰胺跟随了的 HUCBMNC 被对待。在处理和肝织物是免疫组织化学检测的学习的组织病理学说的联盟者和人的法新社和白长袍的以后,老鼠在不同时间点被打死。在肝织物的人的 X 不活跃特定的抄本基因碎片被 PCR 放大发现人的 DNA。结果:房间文化的结果证明支持者房间在控制组是为法新社或白长袍的染色的 negative。然而,在劝诱的组的支持者房间染色了为法新社或白长袍的积极。动物实验的结果显示出那个不人的法新社或在组 A (控制组) 的肝织物在场的白长袍的积极细胞。然而,许多人的法新社或白长袍的积极房间在湾穴附近被散布肝我们和肝的腹片并且在在在一个月以后的组 B 和组 C 的门区域的中央静脉。人的 X 浓度基因的碎片能在组 B 和 C 的肝织物,然而并非在组 A 被检测。结论:在进肝细胞在活体内和在试管内的某些条件 HUCBSC 罐头 differentiate 下面。
AIM: To study the condition and potentiality of human umbilical cord blood stem cells (HUCBSC) to differentiate into hepatocytes in vivo or in vitro.
METHODS: In a cell culture study of human umbilical cord blood stem cell (HUCBSC) differentiation, human umbilical cord blood mononuclear cells (HUCBMNC) were separated by density gradient centrifugation. Fibroblast growth factor (FGF) and hepatocyte growth factor (HGF) and the supernatant of fetal liver were added in the inducing groups. Only FGF was added in the control group. The expansion and differentiation of HUCBMNC in each group were observed. Human alpha fetoprotein (AFP) and albumin (ALB) were detected by immunohistochemistry. In the animal experiments, the survival SD rats with acute hepatic injury after carbon tetrachloride (CCL4) injection 48 h were randomly divided into three groups. The rats in group A were treated with human umbilical cord blood serum. The rats in group B were treated with HUCBMNC transplantation. The rats in group C were treated with HUCBMNC transplantation followed by intraperitoneal cyclophosphamide for 7 d. The rats were killed at different time points after the treatment and the liver tissue was histopathologically studied and human AFP and ALB detected by immunohistochemistry. The human X inactive-specific transcript gene fragment in the liver tissue was amplified by PCR to find human DNA.
RESULTS: The results of cell culture showed that adherent cells were stained negative for AFP or ALB in control group. However, the adherent cells in the inducing groups stained positive for AFP or ALB. The result of animal experiment showed that no human AFP or ALB positive cells present in the liver tissue of group A (control group). However, many human AFP or ALB positive cells were scattered around sinus hepaUcus and the central veins of hepatic Iobules and in the portal area in group B and group C after one month. The fragment of human X chromagene could be detected in the liver tissue of groups B and C, but not in group A.
CONCLUSION: Under certain conditions HUCBSC can differentiate into liver cells in vivo and in vitro.
基金
Supported by the National Natural Science Foundation of China, No. 39870651Natural Science Foundation of Hunan Province, No. 04JJ6048
关键词
脐带血
干细胞
肝细胞
细胞移植
细胞分化
Umbilical cord blood
Stem cell
Liver failure
Cell transplantation
Cell differentiation