Development of cell therapy for hepatic failure due to dysfunction of hepatic parenchymal cells.

开发细胞疗法治疗肝实质细胞功能障碍所致的肝衰竭。

基本信息

批准号：
09470273
负责人：
AMEMIYA Hiroshi
金额：
$ 7.62万
依托单位：
National Children's Medical Research Center
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

Although there are various kinds of novel mdical technology at present, hepatic failure is one of serious diseases which have no fundamental therapies except for liver transplantation or only allopathy such as ultraviolet irradiation against hyperbilirubinemia etc, and some of the hepatic failure are due to congenital malfunction. In this research, we did basic investigation on hepatic cell therapy which may replace liver transplantation or innovate gene therapy. The main topics are 1) search off actor(s) or condition to develop or to maintain differentiated function of hepatocytes, 2) suitable cell types for hepatic cell transplantation, and 3) conditionhg of successful cell transplantation by gene transduction. We developed intra-gel culture of rat hepatocytes with rat fetus extract and rat plasma. This culture condition maintained drug metabolizing activity assessed by 7-ethoxycoumarin deethylation tip to for 7 days, as well as morphological feature of hepatocyte. For cell transplantation, we used amniotic epithelial cells from rat fetus. The certain population of the amniotic cells expressed albumin during culture period as was reported by human amniotic epithelial cells (J Hum Genet 45 171 2000). The cells can be stored frozen without marked impairment of viability. The cell division was 2.7 times at average and telomerase activity was detectable in the cells from feyses at early stage (day 14), suggesting that telomerase transfection may accelerate eell proliferation. To establish successful eell survival after transplantation, ants-apoptotic gene, CrmA, was transfected to hepatocytes, which showed long-term survival against rejection response.

虽然目前的医疗技术多种多样，但肝衰竭是一种严重的疾病，除了肝移植或仅采用紫外线照射对抗高胆红素血症等对抗疗法外，没有根本性的治疗方法，部分肝衰竭是由于先天性功能障碍所致。。在这项研究中，我们对肝细胞疗法进行了基础研究，该疗法可能取代肝移植或创新基因疗法。主要主题是1）寻找发展或维持肝细胞分化功能的因素或条件，2）适合肝细胞移植的细胞类型，以及3）通过基因转导成功进行细胞移植的条件。我们用大鼠胎儿提取物和大鼠血浆开发了大鼠肝细胞的凝胶内培养物。该培养条件维持7-乙氧基香豆素去乙基化尖端评估的药物代谢活性7天，以及肝细胞的形态特征。对于细胞移植，我们使用来自大鼠胎儿的羊膜上皮细胞。正如人羊膜上皮细胞报道的那样，某些羊膜细胞群在培养期间表达白蛋白(J Hum Genet 45 171 2000)。细胞可以冷冻保存而不会明显损害活力。细胞平均分裂2.7次，并且在早期（第14天）细胞中可检测到端粒酶活性，表明端粒酶转染可能加速细胞增殖。为了确保移植后细胞成功存活，将蚂蚁凋亡基因 CrmA 转染至肝细胞，该肝细胞表现出针对排斥反应的长期存活。