Serum-free culture of the MKN-45 gastric cancer cell line and preparation of stem cells
10.16571/j.cnki.1008-8199.2016.04.009
- VernacularTitle:胃癌 MKN-45细胞系的无血清培养技术及干细胞的制备
- Author:
Yiqian JIANG
;
Xiaoxiang GUAN
;
Qingmin GUO
;
Xiaoping XU
;
Suhong AN
;
Yiyang HE
;
Juncai LIANG
;
Lijuan HAN
- Publication Type:Journal Article
- Keywords:
Cancer stem cell;
Serum-free cultivation;
Side population cell;
CDl33 +;
CD44 +
- From:
Journal of Medical Postgraduates
2016;29(4):380-383
- CountryChina
- Language:Chinese
-
Abstract:
[Abstract ] Objective Difficulties with the preparation of gastric cancer stem cells (CSCs) have been a main obstacle to the studies of gastric cancer .This article addresses the technology of the serum-free medium suspension cultivation of the MKN-45 gastric cancer cell line and screening of stem cells from the cell line based on the biomarkers of gastric CSCs . Methods MKN-45 cells were cultured in serum-free medium for 8 weeks and those in the logarithmic phase cultivated with hoechst 33342 followed by detection of the side cells by flow cytometry .When the side population cells reached 25%, all the cell microspheres were collected , hatched with CD133 and CD44, and subjected to fluorescence-activated cell sorting.The CDl33 +and CD44 +cells were selected as gastric CSCs . Results About 40%of the MKN-45 gastric CSCs were alive , prolif-erated, and formed floating cell balls .Side population cells constitu-ted 3.4% of the MKN-45 cells and 26.9% of the cell balls.The CDl33 +and CD44 +cells made up 11.2% of the MKN-45 cells and 90.3%of the cell balls. Conclusion Cell balls rich in CSCs can be successfully obtained by serum-free medium suspension culti-vation and CSCs can be screened out with hoechst 33342 and surface markers , which may serve as an experimental ground for the stud-ies of gastric CSCs .
