RRC ID |
31998
|
著者 |
Yo M, Sakaue-Sawano A, Noda S, Miyawaki A, Miyoshi H.
|
タイトル |
Fucci-guided purification of hematopoietic stem cells with high repopulating activity.
|
ジャーナル |
Biochem Biophys Res Commun
|
Abstract |
Fluorescent ubiquitination-based cell cycle indicator (Fucci) technology utilizing the cell cycle-dependent proteolysis of ubiquitin oscillators enables visualization of cell cycle progression in living cells. The Fucci probe consists of two chimeric fluorescent proteins, FucciS/G2/M and FucciG1, which label the nuclei of cells in S/G2/M phase green and those in G1 phase red, respectively. In this study, we generated Fucci transgenic mice and analyzed transgene expression in hematopoietic cells using flow cytometry. The FucciS/G2/M-#474 and FucciG1-#639 mouse lines exhibited high-level transgene expression in most hematopoietic cell populations. The FucciG1-#610 line expressed the transgene at high levels predominantly in the hematopoietic stem cell (HSC) population. Analysis of the HSC (CD34(-)KSL: CD34(-/low)c-Kit(+)Sca-1(+)lineage marker(-)) population in the transgenic mice expressing both FucciS/G2/M and FucciG1 (#474/#610) confirmed that more than 95% of the cells were in G0/G1 phase, although the FucciG1(red) intensity was heterogeneous. An in vivo competitive repopulation assay revealed that repopulating activity resided largely in the FucciG1(red)(high) fraction of CD34(-)KSL cells. Thus, the CD34(-)KSL HSC population can be further purified on the basis of the Fucci intensity.
|
巻・号 |
457(1)
|
ページ |
7-11
|
公開日 |
2015-1-30
|
DOI |
10.1016/j.bbrc.2014.12.074
|
PII |
S0006-291X(14)02256-6
|
PMID |
25534850
|
MeSH |
Animals
Cell Cycle*
Cell Separation / methods*
Flow Cytometry
Fluorescence
Fluorescent Dyes / metabolism*
Hematopoietic Stem Cells / cytology*
Hematopoietic Stem Cells / metabolism
Mice, Transgenic
Transgenes
Ubiquitination*
|
IF |
2.985
|
引用数 |
7
|
WOS 分野
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
実験動物マウス |
RBRC00144
RBRC02704
RBRC02708
RBRC02817 |