3D culture and analysis of the expression of cancer stem cell markers from gastric cancer cell line MKN45

Abstract

Gastric cancer is the fourth leading cause of worldwide cancer mortality, Vietnam belongs to the specified high-risk group of developing gastric cancer. Gastric cancer stem cells (CSCs) are considered to be the origin of gastric adenocarcinoma. 3D cell culture is an important step of identification of stem cell as well as CSC. In this study, a non-adherent cell culture model (3D) was developed with a culture medium that had growth factors selective for the development of gastric CSCs from gastric cancer cell line MKN45. The result showed that only 6% single cancer cells developed into tumorspheres during culture process. Then, flow cytometry and immunofluorescence analysis was carried out to evaluate the expression of CSC markers CD44 and ALDH in samples taken from tumorspheres which arose from gastric CSCs. The results showed that gastric CSCs formed tumorspheres in 3D culture condition with a high expression of 92% and 40,1% for markers CD44 and ALDH, respectively.