Musashi1 expression cells derived from mouse embryonic stem cells can be enriched in side population isolated by fluorescence activated cell sorter.
BACKGROUND
Purifying stem cells is an inevitable process for further investigation and cell-therapy. Sorting side population (SP) cells is generally regarded as an effective method to enrich for progenitor cells. This study was to explore whether sorting SP could enrich for the Musashi1 (Msi1) positive cells from Msi1 high expression cells (Msi1(high) cells) derived from mouse embryonic stem cells (ESCs) in vitro.


RESULTS
In this study, Msi1(high) cell population derived from ESCs were stained by Hoechst 33342, and then the SP and non-SP (NSP) fractions were analyzed and sorted by fluorescence activated cell sorter. Subsequently, the expressions of Msi1 and other markers for neural and intestinal stem cells in SP and NSP were respectively detected. SP and NSP cells were hypodermically engrafted into the backs of NOD/SCID mice to form grafts. The developments of neural and intestinal epithelial cells in these grafts were investigated. SP fraction was identified and isolated from Msi1(high) cell population. The expression of Msi1 in SP fraction was significantly higher than that in NSP fraction and unsorted Msi1(high) cells (P< 0.05). Furthermore, the markers for neural cells and intestinal epithelial cells were more highly expressed in the grafts from SP fraction than those from NSP fraction (P< 0.05).


CONCLUSIONS
SP fraction, isolated from Msi1(high) cells, contains almost all the Msi1-positive cells and has the potential to differentiate into neural and intestinal epithelial cells in vivo. Sorting SP fraction could be a convenient and practical method to enrich for Msi1-positive cells from the differentiated cell population derived from ESCs.
Department of Gastroenterology, the Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, People's Republic of China.
Tao
T
Yu
Li-Na
LN
Zhao
Shao-Yang
SY
Lan
Miao-Jing
MJ
Fan
Yu
Y
Gong
Liu
L
Shi
Yu-Hong
YH
Yuan
Kai-Hong
KH
Huang
Qi-Kui
QK
Chen
D000818:Animals; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D051379:Mice; D016513:Mice, SCID; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000032:analysis* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
C104786:Msi1h protein, mouse; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins