TY - JOUR
T1 - Messenger RNA quantification after fluorescence activated cell sorting using intracellular antigens
AU - Yamada, Hiroya
AU - Maruo, Rie
AU - Watanabe, Mikio
AU - Hidaka, Yoh
AU - Iwatani, Yoshinori
AU - Takano, Toru
N1 - Funding Information:
This research was supported by the Ministry of Education, Culture, Sports, Science, and Technology of Japan via a Grant-in-Aid for Scientific Research C, 2008–2010, No. 20590570 ; a Research Grant from the Princess Takamatsu Cancer Research Fund 04-23606 ; and the Japanese Society of Laboratory Medicine Fund for the Promotion of Scientific Research .
PY - 2010/7
Y1 - 2010/7
N2 - Recent studies using stem cells or cancer stem cells have revealed the importance of detecting minor populations of cells in blood or tissue and analyzing their biological characteristics. The only possible method for carrying out such procedures is fluorescence activated cell sorting (FACS). However, FACS has the following limitations. First, cells without an appropriate cell surface marker cannot be sorted. Second, the cells have to be kept alive during the sorting process in order to analyze their biological characteristics. If an intracellular antigen that was specific to a particular cell type could be stained with a florescent dye and then the cells can be sorted without causing RNA degradation, a more simple and universal method for sorting and analyzing cells with a specific gene expression pattern could be established since the biological characteristics of the sorted cells could then be determined by analyzing their gene expression profile. In this study, we established a basic protocol for messenger RNA quantification after FACS (FACS-mQ) targeting intracellular antigens. This method can be used for the detection and analysis of stem cells or cancer stem cells in various tissues.
AB - Recent studies using stem cells or cancer stem cells have revealed the importance of detecting minor populations of cells in blood or tissue and analyzing their biological characteristics. The only possible method for carrying out such procedures is fluorescence activated cell sorting (FACS). However, FACS has the following limitations. First, cells without an appropriate cell surface marker cannot be sorted. Second, the cells have to be kept alive during the sorting process in order to analyze their biological characteristics. If an intracellular antigen that was specific to a particular cell type could be stained with a florescent dye and then the cells can be sorted without causing RNA degradation, a more simple and universal method for sorting and analyzing cells with a specific gene expression pattern could be established since the biological characteristics of the sorted cells could then be determined by analyzing their gene expression profile. In this study, we established a basic protocol for messenger RNA quantification after FACS (FACS-mQ) targeting intracellular antigens. This method can be used for the detection and analysis of stem cells or cancer stem cells in various tissues.
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U2 - 10.1016/j.bbrc.2010.05.112
DO - 10.1016/j.bbrc.2010.05.112
M3 - Article
C2 - 20510885
AN - SCOPUS:77954218683
SN - 0006-291X
VL - 397
SP - 425
EP - 428
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -