Benoist S, Sarkis R, Barbu V, Honiger J, Baudrimont M, Lakehal F, Becquemont L, Delelo R, Housset C, Balladur P, Capeau J, Nordlinger B
Research Unit 402 of INSERM, Paris, France.
Surgery. 2001 May;129(5):606-16. doi: 10.1067/msy.2001.112961.
This study evaluated the survival and functions of encapsulated porcine hepatocytes after intraperitoneal allotransplantation and xenotransplantation without immunosuppression.
Isolated porcine hepatocytes were encapsulated in AN 69 polymer capsules (45.10(6)/capsule) and transplanted intraperitoneally in 12 rats and 12 pigs. Fifteen, 30, and 60 days after transplantation, capsules were removed and the viability and morphology of explanted hepatocytes were examined under light and electronic microscopy. The potential to produce albumin was assessed by evaluating the level of albumin messenger RNA, using semiquantitative reverse transcription-polymerase chain reaction. 6beta-Hydroxylase activity was measured by high-performance liquid chromatography. In addition, cytochrome P450 3A proteins were detected by Western blot only in allogeneic hepatocytes.
Similar results were observed after allotransplantation and xenotransplantation. Histologic studies showed that hepatocytes were well-preserved and arranged in cords for up to 30 days. The expression of porcine albumin gene was maintained up to 15 days. 6beta-Hydroxylase activity was 2.5-fold lower at day 15 than in freshly encapsulated hepatocytes, which were not transplanted. In allogeneic hepatocytes, the expression of CYP 3A protein was detected up to 60 days after transplantation.
Encapsulated porcine hepatocytes remain viable and functional for at least 15 days after allotransplantation and xenotransplantation without immunosuppression. The demonstration of maintained hepatic functions in transplanted porcine hepatocytes up to 15 days is a first step toward application in the treatment of acute liver failure.
本研究评估了在无免疫抑制情况下，经腹腔同种异体移植和异种移植后，包封猪肝细胞的存活情况及功能。
将分离的猪肝细胞包封于AN 69聚合物胶囊中（每个胶囊45×10⁶个细胞），并经腹腔移植到12只大鼠和12只猪体内。移植后15天、30天和60天，取出胶囊，在光学显微镜和电子显微镜下检查移出肝细胞的活力和形态。通过使用半定量逆转录-聚合酶链反应评估白蛋白信使核糖核酸水平来评估产生白蛋白的潜力。采用高效液相色谱法测量6β-羟化酶活性。此外，仅在同种异体肝细胞中通过蛋白质印迹法检测细胞色素P450 3A蛋白。
同种异体移植和异种移植后观察到相似结果。组织学研究表明，肝细胞保存良好，呈索状排列长达30天。猪白蛋白基因的表达维持长达15天。与未移植的新鲜包封肝细胞相比，15天时6β-羟化酶活性降低了2.5倍。在同种异体肝细胞中，移植后60天内均可检测到CYP ³A蛋白的表达。
在无免疫抑制的情况下，经腹腔同种异体移植和异种移植后，包封猪肝细胞至少15天内仍保持存活且具有功能。移植猪肝细胞中肝功能维持长达15天的这一证明是朝着应用于急性肝衰竭治疗迈出的第一步。
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