Abstract
BackgroundPurifying 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 (Msi1high cells) derived from mouse embryonic stem cells (ESCs) in vitro.ResultsIn this study, Msi1high 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 Msi1high cell population. The expression of Msi1 in SP fraction was significantly higher than that in NSP fraction and unsorted Msi1high 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).ConclusionsSP fraction, isolated from Msi1high 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.
Highlights
Purifying stem cells is an inevitable process for further investigation and cell-therapy
We had found that Msi1 and hairy and enhancer of split 1 (Hes1) high-expression cells (Msi1highHes1high cells) derived from mouse embryonic stem cells (ESCs) could develop into small intestinal epithelial cells, which enhanced the repair of small intestinal injury in vivo [23]
Differentiation of Msi1high cells in vitro ESCs were cultured by the hanging-drop method in embryonic bodies (EBs) medium to attain EBs in vitro
Summary
Purifying stem cells is an inevitable process for further investigation and cell-therapy. This study was to explore whether sorting SP could enrich for the Musashi (Msi1) positive cells from Msi high expression cells (Msi1high cells) derived from mouse embryonic stem cells (ESCs) in vitro. 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 Msi1high 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. Embryonic stem cells (ESCs) are pluripotent cells derived from the inner cell mass of the mammalian blastocyst with self-renewal capacity and multi-developmental plasticity, which makes ESCs a powerful tool for cell-based therapy [1,2]. A more convenient and practical separation strategy should be established for the further investigation of Msi1-positive cells
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