Modest isolation and enrichment of intestinal epithelial stem cells by rhodamine 123 Staining
10.3760/cma.j.issn.1673-4203.2010.09.007
- VernacularTitle:利用Rhodamine 123染色初步分离及纯化肠上皮干细胞
- Author:
Yuqi LUO
;
Chengtang WU
;
Weishan CAI
;
Zhan PENG
- Publication Type:Journal Article
- Keywords:
Intestinal epithelial stem cells;
Rhodamine 123;
Musashi-1
- From:
International Journal of Surgery
2010;37(9):596-599
- CountryChina
- Language:Chinese
-
Abstract:
Objective To discuss the technology of modest isolating and enriching the intestinal epithelial stem cells. Methods Mouse intestinal epithelial cells were stained by Rhodamine 123 (Rho), sorting the Rhodamine 123 low staining cell population ( Rholow ) and Rhodamine 123 strong straining cell population ( Rhobri ) by fluorescence activated cell sorting(FACS) in flow cytometer; Detecting the musaashi-1 and p-glycoprotein 1 (p-g-p1) mRNA expression of two groups by RT-PCR; Analyzing the cell cycle and the percentage of the musashi-1 positive cells by flow cytometry. Results The intestinal epithelial cells were divided into three groups, Rhodamine 123 low staining cells( 12. 34% of total cells), Rhodamine 123 middle staining cells (45.26% of total cells) and Rhodamine 123 strong staining cells ( 41. 40% of total cells). The Rholow cell fraction and Rhobri cell fraction were isolated successfully. Both of musashi-1 and p-g-p1 mRNA were strongly expressed in Rho1ow cell fraction, and Rhobri cell fraction little expressed p-g-p1 mRNA. Most of Rho1ow cells were in G0/G1 phase, and the musashi-1 positive cells were about 10.37% of total cells in this fraction. Conclusion The intestinal epithelial stem cells can be modestly isolated and enriched by Rhodamine 123 staining.
