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将小鼠随机分为治疗组和对照组,按20μL/g的剂量腹腔注射10%CCl4溶液,每周2次,连续注射4周,建立小鼠急性肝损伤模型。建模后24h,治疗组尾静脉移植经Dir标记的人羊膜间充质干细胞0.2mL;对照组移植等量未经Dir标记的人羊膜间充质干细胞。使用小动物活体成像系统观察人羊膜间充质干细胞在小鼠体内分布。将NOD/SCID小鼠随机分为3组,试验组注射0.2 mL人羊膜间充质干细胞细胞悬液,阳性对照组注射0.2 mL Hep G2细胞悬液,阴性对照组只注射等量Matrigel和PBS的1∶1混合液。60d后脱颈处死小鼠,观察肿瘤形成情况并进行病理组织学检查。结果显示,活体成像试验发现,人羊膜间充质干细胞移植后2h,全身只有肝脏区域有荧光,人羊膜间充质干细胞移植后1d,肝脏及肺脏附近荧光信号达到最强,移植3d,肺部强荧光消失,肝部附近强荧光仍存在,移植后7~30d,全身各处荧光信号消失,但肝部荧光信号持续存在。体内致瘤试验发现,阳性对照组小鼠有肿瘤形成,而注射人羊膜间充质干细胞的试验组小鼠和阴性对照组小鼠均未观察到肿瘤形成。结果表明,人羊膜间充质干细胞尾静脉移植入急性肝损伤小鼠,可以成功归巢至肝脏并长期定植存在;人羊膜间充质干细胞在体内没有致瘤性,具有一定的临床安全性。
The mice were randomly divided into treatment group and control group. Acute liver injury model was established by intraperitoneal injection of 10% CCl4 solution at the dose of 20 μL / g twice a week for 4 weeks. After modeling 24 h, the tail vein of the treatment group was transplanted with 0.2 mL of Dir-labeled human amniotic mesenchymal stem cells; the control group was transplanted with an equal number of human amnion-derived mesenchymal stem cells without Dir. Small animal live imaging system was used to observe the distribution of human amniotic mesenchymal stem cells in mice. The NOD / SCID mice were randomly divided into 3 groups: 0.2 mL human amniotic mesenchymal stem cell suspension was injected into the experimental group, 0.2 mL Hep G2 cell suspension was injected into the positive control group, and only the same amount of Matrigel and PBS were injected into the negative control group 1: 1 mixture. After 60 days, mice were killed by atrophy and observed for tumor formation and histopathological examination. The results showed that only 2 hours after human amniotic mesenchymal stem cell transplantation, the whole body had fluorescence only in the liver region. The fluorescence signal of the liver and lung reached the strongest one day after transplantation of human amniotic mesenchymal stem cells, Strong fluorescence disappeared, strong fluorescence around the liver still exists, 7 ~ 30d after transplantation, the whole body fluorescence signal disappeared, but the liver fluorescence signal persisted. In vivo tumorigenicity test found that the positive control group mice had tumor formation, and injection of human amniotic mesenchymal stem cells in the experimental group and the negative control group mice were not observed tumor formation. The results showed that human amniotic mesenchymal stem cells transplanted into the tail vein of acute liver injury mice successfully homing to the liver and long-term colonization exist; human amniotic mesenchymal stem cells in the body is not tumorigenic, with a certain clinical safety.