Consideration of a Safe Protocol for Hepatocyte Transplantation Using Infantile Pigs
Consideration of a Safe Protocol for Hepatocyte Transplantation Using Infantile Pigs
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DOI：
10.3727/215517912x639469
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Kasahara, Mureo
中科院分区：
文献类型：
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作者：
Enosawa, Shin;Yuan, Wenji;Kasahara, Mureo
肝细胞移植有望成为某些肝移植病例的替代治疗方法。最有希望的适应症是先天性代谢疾病，尤其是婴儿。为了建立婴儿肝细胞移植的安全方案，我们检查了幼猪治疗期间的生理变化。受体猪（家养猪，与大约克夏猪、长白猪和杜洛克猪杂交；2.5千克；7-14天龄）用异氟烷麻醉；最小长度的中线切口提供了进入肠系膜上静脉的通道。将双腔导管通过静脉插入肝门区 1 cm 以内。监测心率、全身血压和门静脉压力等生理参数。从同一品系中分离出的冷冻保存的猪肝细胞悬浮于生理盐水中并通过导管输注。在需要追踪的实验中，细胞在输血前用荧光染料染色。将受体猪饲养1天并处死以进行肝脏组织学检查。经过初步实验，确定肝细胞移植的最佳数量和浓度为1×10(8)cells/kg和1×10(7)cells/ml。以0.67ml/min的速率输注细胞悬浮液。没有观察到明显的生理参数异常，而在初步试验中，当输血速度快于我们的标准方案时，出现了轻微的心动过速。采用既定方法输血的所有五头猪均从麻醉中苏醒，并存活并具有良好的生命体征。组织学显示，输注的肝细胞整合到肝组织中。
Hepatocyte transplantation is hoped to be an alternative treatment to certain cases of liver transplantation. The most promising indication is for congenital metabolic diseases, especially in infants. To establish a safe protocol for hepatocyte transplantation into infants, we examined physiological changes during treatment using infantile pigs. Recipient pigs (domestic, crossbred with Large-Yorkshire, Landrace, and Duroc; 2.5 kg; 7-14 days old) were anesthetized with isoflurane; a midline incision of minimum length provided access to the superior mesenteric vein. A double lumen catheter was inserted through the vein to within 1 cm of the hepatic portal region. Physiological parameters such as heart rate, systemic blood pressure, and portal pressure were monitored. Cryopreserved porcine hepatocytes isolated from the same strain were suspended in physiological saline and transfused through the catheter. In experiments requiring tracing, cells were stained with fluorescent dye prior to transfusion. Recipient pigs were kept for 1 day and sacrificed for histological liver examination. After preliminary experiments, the optimized number and concentration for hepatocyte transplantation were determined to be 1 x 10(8) cells/kg and 1 x 10(7) cells/ml. The cell suspension was transfused at a rate of 0.67 ml/min. No marked anomaly of physiological parameters was observed, whereas light tachycardia occurred in preliminary trials when the transfusion rate was faster than our standard protocol. All five pigs transfused with the established method recovered from anesthesia and survived with good vital signs. Histology revealed that the transfused hepatocytes were integrated in the hepatic tissue.