In recent years, chemotherapy targeting cancer stem cells has increasingly garnered interest from the scientists and clinicians. In this study, we explored both in vitro and in vivo if a chemotherapy regimen could arrest gastric tumor growth via demolishing CD24+ and CD44+ cancer stem cells (CSC) in the tumor mass. To facilitate the tumorigenesis in the subcutaneous xenograft model, we employed an electrospun scaffold comprising polydioxanone and gelatin for the delivery of tumor cells. The scaffold sported a highly porous micro-structure and promoted the in vitro and in vivo tumorigenesis without inducing apoptosis. in vivo studies confirmed that the tumor enjoyed an increase of volume by 56.21% and weight by 71.29%, respectively, in a 12-week period. Interestingly, populations of CD24+ and CD44+ cells in the tumor increased by 37% and 274%, respectively. No phenotypic change during tumorigenesis was observed due to the presence of the scaffold. Thereafter, we studied the chemotherapeutic effectiveness of a regimen comprising docetaxel, cisplatin and fluorouracil (DCF) in the scaffold model. The study showed that this regimen demonstrated a strong capability to demolish the CD44+ subpopulation in the tumor. This discovery warrants further study of the DCF regimen for gastric cancer. Moreover, the scaffold offers cancer scientists an accelerated in vivo platform for mechanistic chemotherapy study.