Expansion of human hematopoiteic stem cells in transgenic or large animals.

在转基因动物或大型动物中扩增人类造血干细胞。

基本信息

批准号：
07557077
负责人：
NAKAUCHI Hiromitsu
金额：
$ 10.24万
依托单位：
University of Tsukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557077/
关键词：
Hematopoietic stem cells GM-CSF IL-3 swine transgenic mice SCID mice Myelodysplastic syndrome NK cells.自己複製サイトカイン G-CSF GM- CSF

项目摘要

1.Engraftment of human hematopoietic cells in transgenic Severe Combined Immunodeficient mice.A transgenic SCID (TG-SCID) mouse expressing the human cytokines interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) has been generated with the aim of making a model system allowing the in vivo proliferation of human hematopoietic cells. Using TG-SCID mice expressing high levels (30-35 ng/ml in the serum) of human GM-CSF and IL-3, we attempted to engraft a human myeloid leukemia cell line, F-36P,derived from a myelodysplastic syndrome (MDS) patient. When F-36P cells were transferred intravenously into sublethally irradiated TG-SCID mice, extensive proliferation of F-36P cells was found four to six weeks later. Successful engraftment, however, required the pre-administration of a monoclonal antibody to mouse interleukin-2 receptor (IL-2R) beta chain, neutralizing NK activity. Surprisingly, all of the transplanted TG-SCID mice engrafted with F-36P cells developed … More hind leg paralysis approximately 6 weeks after transfer. Histological analysis demonstrated extensive invasion and formation of osteolytic lesions by the F-36P cells in the vertebrae. These data indicate that transgenic SCID mice expressing human IL-3 and GM-CSF provide a useful system for the study of human leukemia. In addition, NK cells appear to play an important role in rejection of human cells.2.Transplantation of human hematopoietic cells into miniature-swine and swine fetusTo establish a model for a long-term human hematopoietic reconstitution, human bone marrow or cord blood cells were transplanted into day 45-81 miniature swine or swine fetuses through uterine wall of a pregnant mother. Of the 15 pregnant animals to which transplantation of human hematopoietic cells was performed, five of them gave birth. New born pigs were examined periodically for the presence of human CD45_+ blood cells in peripheral blood and in bone marrow. So far, none of them have human CD45_+ cells above 1% level either in PB or in bone marrow. Less

1.转基因严重的免疫缺陷小鼠中人类造血细胞的过育。表达人细胞因子介菌素三（IL-3）（IL-3）和粒细胞 - 摩托噬细胞刺激因子（GM-CSF）的人性细胞因子（GM-CSF）的刺激性范围的人类细胞因子的刺激剂量已成为模型的瞄准剂，这表达了人细胞因子Iltleukin-3（IL-3）和粒细胞噬菌体刺激因子（IL-3）和粒细胞 - 摩托结构刺激因子（IL-3）和粒细胞刺激性刺激因子（GM-CSF）细胞。使用人GM-CSF和IL-3的高水平（30-35 ng/mL）的TG-SCID小鼠，我们试图植入源自骨髓质量异型综合征（MDS）患者的人髓性白血病细胞系F-36P。当将F-36P细胞静脉注射转移到量照射的TG-SCID小鼠中时，在四到六周后发现了F-36p细胞的广泛增殖。然而，成功的植入需要对小鼠白介素-2受体（IL-2R）β链的单克隆抗体进行预先施用，从而中和NK活性。出人意料的是，所有被F-36P细胞烧伤的移植的TG-SCID小鼠……在转移后大约6周后，更多的后腿瘫痪。组织学分析表明，椎骨中F-36p细胞对骨化病变的广泛侵袭和形成。这些数据表明表达人IL-3和GM-CSF的转基因SCID小鼠为研究人白血病提供了有用的系统。 In addition, NK cells appear to play an important role in rejection of human cells.2.Transplantation of human hematopoietic cells into miniature-swine and swine fetusTo establish a model for a long-term human hematopoietic reconstitution, human bone marrow or cord blood cells were transplanted into day 45-81 miniature swine or swine fetuses through uterine wall of a pregnant mother.在进行人类造血细胞移植的15只孕妇中，其中5只生了。定期检查新生的猪是否存在外周血和骨髓中人类CD45_+血细胞的存在。到目前为止，在PB或骨髓中，它们都没有人类CD45_+细胞高于1％的水平。较少的