Characterization of the hematopoietic stem cell and search for the molecules responsible for stem cell self-renewal.

造血干细胞的表征并寻找负责干细胞自我更新的分子。

基本信息

批准号：
08407028
负责人：
NAKAUCHI Hiromitsu
金额：
$ 22.34万
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：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08407028/
关键词：
hematopoietic stem cell CD34 hematopoietic Progenitor cells cell cycle lineage commitment SCF G-CSF mobilization 末梢血幹細胞移植骨髄移植自己複製能多分化能 paired-daughter cells Stochastic model seeding efficiency 長期骨髄再建能

项目摘要

1) We have determinedthe phenotype of hematopoietic stem cells (HSC) and hematopoietic progenitor cells in adult mouse bone marrow. They were CD34^-c-Kit^+Sca-1^+ Lineage marker(Lin)^- and CD34^+ c-Kit^+Sca-1^+Lin^-, respectively. By combining these two populations, we have developeda sensitive stem cell assay nameda progenitor rescue assay. Using this assay, we were able to show a single HSC can reconstitute bone marrow of a lethally irradiated mouse for over 300 days.2) In 6 week-old mice, CD34^-c-Kit^+ Sca-1^+Li^- HSC can be found at a frequency of 1 in 25000-20000 bone marrow cells. However, in older mice, both relative frequency and absolute number of HSC were increased significantly. HPP-CFU as well as long-term marrow repopulation assays also supported this finding.3) Propidium iodide staining of the purified HSC revealedthat over 97% of them were in G0G1 phase. Long-termBrdU uptake experiment showed that over 70% of HSC take up BrdU within the first two weeks. These data indicate that majority of HSC contribute to hematopoiesis but they devide rather infrequently.4) Paired-daughtercell experiment demonstratedthat commitment of HSC appear to be stochastic although among paired-daughtercolonies, cells of certain lineages appeared significantly higher incidence suggesting non-randomfashion of the lineage commitment.5) We happenedto find out that mouse HSC are CD34-low/negative. It is thereforecritically important to examine if there are CD34-low/negative HSC in human. Establishment of a solid assay system for humanHSC is necessary.

1）我们确定了成年小鼠骨髓中造血干细胞（HSC）和造血祖细胞的表型。它们分别是CD34^-c-Kit^+Sca-1^+谱系标记(Lin)^-和CD34^+c-Kit^+Sca-1^+Lin^-。通过结合这两个群体，我们开发了一种敏感的干细胞测定，称为祖细胞救援测定。通过该测定，我们能够证明单个 HSC 可以在 300 多天内重建受到致命辐射的小鼠的骨髓。2) 在 6 周大的小鼠中，CD34^-c-Kit^+ Sca-1^+Li^ - HSC 的发现频率为 25000-20000 个骨髓细胞中 1 个。然而，在老年小鼠中，HSC 的相对频率和绝对数量均显着增加。 HPP-CFU 以及长期骨髓再增殖测定也支持这一发现。3) 纯化的 HSC 的碘化丙啶染色显示，其中 97% 以上处于 G0G1 期。长期BrdU摄取实验表明，超过70%的HSC在前两周内摄取了BrdU。这些数据表明，大多数 HSC 有助于造血，但它们分裂的频率相当低。 4) 配对子细胞实验表明，HSC 的承诺似乎是随机的，尽管在配对子细胞集落中，某些谱系的细胞出现率显着较高，表明 HSC 的非随机方式。谱系承诺。5) 我们碰巧发现小鼠 HSC 是 CD34 低/阴性。因此，检查人体内是否存在CD34低/阴性的HSC至关重要。建立人类HSC的固体检测系统是必要的。