Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo.
Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo.
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DOI：
10.1084/jem.183.4.1797
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发表时间：
1996-04-01
期刊：
影响因子：
--
通讯作者：
Mulligan RC
中科院分区：
文献类型：
--
作者：
Goodell MA;Brose K;Paradis G;Conner AS;Mulligan RC
造血干细胞(HSC)是一种多能细胞，存在于骨髓中，在动物一生中补充所有成人的造血血统。在用活性染料Hoechst 33342对小鼠骨髓细胞进行染色的实验中，我们发现在两个发射波长同时显示Hoechst荧光显示了一小部分具有多潜能造血干细胞表型标记的完整骨髓细胞。这些细胞在竞争性再繁殖实验中被证明包含来自小鼠骨髓的绝大多数HSC活性，并为体内重建活性至少浓缩了1000倍。此外，这些Hoechst染色的侧群(SP)细胞被证明在低细胞剂量下保护受者免受致命性照射，并对淋巴系和髓系都有贡献。在维拉帕米存在下进行染色时，Hoechst SP轮廓的形成被阻断，表明SP细胞的明显低染色模式是由于多药耐药蛋白(MDR)或MDR样介导的染料从HSC外流所致。阻断Hoechst外流活动的能力也使我们能够使用Hoechst来确定SP细胞的DNA含量。S-G2M的HSC占1%-3%。这也使得纯化的G0-G1和S-G2M造血干细胞具有相当于静止干细胞的重建能力。这些发现对造血细胞发育的模型和涉及造血细胞的疾病的遗传疗法的发展具有重要意义。
Hematopoietic stem cells (HSC) are multipotent cells that reside in the bone marrow and replenish all adult hematopoietic lineages throughout the lifetime of the animal. While experimenting with staining of murine bone marrow cells with the vital dye, Hoechst 33342, we discovered that display of Hoechst fluorescence simultaneously at two emission wavelengths revealed a small and distinct subset of whole bone marrow cells that had phenotypic markers of multipotential HSC. These cells were shown in competitive repopulation experiments to contain the vast majority of HSC activity from murine bone marrow and to be enriched at least 1,000-fold for in vivo reconstitution activity. Further, these Hoechst-stained side population (SP) cells were shown to protect recipients from lethal irradiation at low cell doses, and to contribute to both lymphoid and myeloid lineages. The formation of the Hoechst SP profile was blocked when staining was performed in the presence of verapamil, indicating that the distinctly low staining pattern of the SP cells is due to a multidrug resistance protein (mdr) or mdr-like mediated efflux of the dye from HSC. The ability to block the Hoechst efflux activity also allowed us to use Hoechst to determine the DNA content of the SP cells. Between 1 and 3% of the HSC were shown to be in S-G2M. This also enabled the purification of the G0-G1 and S-G2M HSC had a reconstitution capacity equivalent to quiescent stem cells. These findings have implications for models of hematopoietic cell development and for the development of genetic therapies for diseases involving hematopoietic cells.
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影响因子：
3.2
作者：
SCHEID, MP;TRIGLIA, D
通讯作者：
TRIGLIA, D
DOI：
10.1073/pnas.85.3.822
发表时间：
1988-02-01
影响因子：
11.1
作者：
HARRISON, DE;ASTLE, CM;LERNER, C
通讯作者：
LERNER, C
影响因子：
--
作者：
SIMINOVITCH, L;TILL, JE;MCCULLOCH, EA
通讯作者：
MCCULLOCH, EA
影响因子：
20.3
作者：
SPANGRUDE, GJ;BROOKS, DM;TUMAS, DB
通讯作者：
TUMAS, DB
影响因子：
15.3
作者：
ROSS, EAM;ANDERSON, N;MICKLEM, HS
通讯作者：
MICKLEM, HS