An efficient method for single hematopoietic stem cell engraftment in mice based on cell-cycle dormancy of hematopoietic stem cells.
An efficient method for single hematopoietic stem cell engraftment in mice based on cell-cycle dormancy of hematopoietic stem cells.
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DOI：
10.1016/j.exphem.2010.03.013
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发表时间：
2010-07
影响因子：
2.6
通讯作者：
Ogawa, Makio
中科院分区：
文献类型：
--
作者：
Abe, Takanori;Masuya, Masahiro;Ogawa, Makio
目的：研究一种高效的单造血干细胞（HSC）移植方法，用于高水平造血移植。我们将单细胞分选与推测的造血干细胞短期培养相结合。将高富集hsc的小鼠骨髓细胞单独置于96孔培养板中，在小鼠c-kit配体和小鼠白细胞介素-11或人重组粒细胞集落刺激因子的存在下孵育。一周后，产生的细胞克隆被单独移植到受到致命辐射的受体中。我们还对单个克隆的增殖进行了时间过程分析。最后，我们对单细胞的配对后代进行了显微处理，并再次研究了造血干细胞结合移植的自我更新和分化潜力。培养第7天的无性系大小与移植后2个月的移植物大小存在相关性。小型克隆，例如由少于15个细胞组成的克隆，通常显示出高水平的多系移植，而由40个或更多细胞组成的克隆显示出非常低水平的移植。对单个克隆细胞分裂的日常观察显示，尽管存在允许细胞因子，一些造血干细胞仍处于G0状态长达1周。对配对后代进行显微操作的研究证明了造血干细胞产生两个造血干细胞的能力以及不对称细胞分裂的能力。单细胞分选结合单个推定造血干细胞的短期培养为单个造血干细胞移植提供了一种有效的方法。单个HSC的动力学分析为HSC细胞周期休眠、自我更新和扩增提供了直接证据。
To develop an efficient method for single hematopoietic stem cell (HSC) transplantation for high-level hematopoietic engraftment. We combined single cell sorting with short-term culture of putative HSCs. Mouse bone marrow cells that had been highly enriched for HSCs were individually deposited into 96 well culture plate and incubated in the presence of mouse c-kit ligand and either mouse interleukin-11 or human recombinant granulocyte colony-stimulating factor. One week later, the resulting clones of cells were individually transplanted into lethally irradiated recipients. We also carried out time course analysis of proliferation of the individual clones. Finally, we used micromanipulation of the paired progenies of the single cells and studied self-renewal and differentiation potentials of HSCs again in combination with transplantation. There was a correlation between clone size at day 7 of culture and engraftment at two months post-transplantation. Small clones, such as those consisting of fewer than 15 cells, often showed high-level multi-lineage engraftment while clones consisting of 40 or more cells showed very low levels of engraftment. Daily observation of cell divisions of individual clones revealed that some HSCs are the in G0 state for as long as 1 week despite the presence of permissive cytokines. Studies using micromanipulation of paired progenies documented the ability of an HSC to generate two HSCs as well as asymmetric cell divisions. Single cell sorting combined with short-term culture of individual putative HSCs provides an efficient method for single HSC transplantation. Analyses of the kinetics of individual HSCs provided direct evidence for HSC cell cycle dormancy, self-renewal and expansion.
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DOI：
10.1073/pnas.80.21.6689
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
SUDA, T;SUDA, J;OGAWA, M
通讯作者：
OGAWA, M
影响因子：
9.2
作者：
Iscove, NN;Nawa, K
通讯作者：
Nawa, K
影响因子：
2.6
作者：
Ebihara, Y;Masuya, M;Ogawa, M
通讯作者：
Ogawa, M
DOI：
10.1073/pnas.84.24.9035
发表时间：
1987-12-01
影响因子：
11.1
作者：
IKEBUCHI, K;WONG, GG;OGAWA, M
通讯作者：
OGAWA, M
DOI：
10.1073/pnas.81.8.2520
发表时间：
1984-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
SUDA, T;SUDA, J;OGAWA, M
通讯作者：
OGAWA, M