EFFECT OF HEPATOCELLULAR TRANSPLANTATION VIA UMBICAL VEIN IN FETAL RATS OF CONGENITAL METABOLIC DISEASE

在亲代期发生和发展的遗传性肝酶缺乏症需要产前治疗。我们研究了通过脐静脉输注肝细胞进行产前肝细胞移植的效果，实验对象是由于肝微粒体缺乏l -古罗内酯氧化酶而无法在肝脏合成抗坏血酸（AsA）的突变体ODS-^<od>/od大鼠。我们还研究了胚胎肝细胞同步移植到成人SHR脾脏后细胞色素P450的表达。1：从ODS-^+/+大鼠肝细胞中分离出与ODS-^<od>/od同源但能合成AsA的肝细胞。雌性ODS-^<od b> /od大鼠妊娠18 d开腹，将其中一胎上方子宫壁固定于腹壁。在子宫壁和羊膜上切开小切口，抽出脐带，显微镜下用30号针注入2-4x10^5个肝细胞至脐静脉。术后6周，给大鼠喂食含有…a的标准饮食。在6只出生存活6周的受体中，有2只大鼠在停止给药后出现发病晚，存活时间是对照组的两倍。这种效应被认为是由植入受体肝脏的肝细胞引起的。II：在妊娠20天从自发性高血压大鼠（SHR）中获取胎肝细胞，然后移植到受体SHR成人脾脏中。受体脾脏形态学检查显示，6周后红髓内可见大块肝细胞，呈明显的索状结构。最重要的是，移植脾脏的肝细胞在移植后2-10周能够表达几个细胞色素P450蛋白家族。因此，移植的肝细胞组织似乎生长并形成细胞色素P450代谢系统。如果在产前诱导免疫耐受，这种治疗可能对重复异体肝细胞移植有很大的优势。少

Prenatal treatment is needed for inherited liver enzyme deficiency which occurs and progresses at parental period. We studied effects of prenatal hepatocellular transplantation by infusion of hepatocytes via the umbilical vein, using ODS-^<od>/od rat, which is a mutant unable to synthesize ascorbic acid (AsA) in the liver because of a lack of L-gulonolactone oxidase in the liver microsomes. And also we studied the expression of cytochromes P450 of fetal hepatocyte transplanted into adult SHR spleen syngenitically. I : Hepatocytes were isolated from ODS-^+/+ rat, which is congeneic to ODS-^<od>/od but can synthesize AsA.Laparotomy was performed on female ODS-^<od>/od rats at 18 days of gestation, and uterus wall above one of the fetuses was fixed to abdominal wall. After small incision was added to the uterus wall and anmion, umbilical cord was drawn out and 2-4x10^5 hepatocytes were infused to the umbilical vein by 30 gauge needle under microscopy. Rats were fed a standard diet contain … More ing AsA for 6 weeks after operation. Out of 6 recipients which were born alive and survived for 6 weeks, 2 rats showed late onset of disease and survived twice as long as controls after the cessation of AsA administration. This effect is thought to be derived from hepatocytes implanted in the liver of recipient. II : Fetal hepatocytes were harvested at 20 days of gestation from spontaneously hypertensive rats (SHR) and then transplanted into recipient adult SHR spleens. Morphological examination of the recipient spleens revealed that after 6 weeks, large masses of hepatocytes were present in the red pulp with apparent cord-like structures. Of major significance was the fact that hepatocyte transplanted spleens were able to express several families of cytochrome P450 proteins 2-10 weeks after transplantation. Thus, the transplanted hepatocyte tissue appeared to grow and develop a cytochrome P450 metabolic system. This therapy might be of great advantage for repeated allogeneic hepatocyte transplantation, if immunological tolerance is induced at prenatal period. Less