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. 2023 May 20;114(8):3259–3269. doi: 10.1111/cas.15840

FIGURE 2.

FIGURE 2

Establishment of patient‐derived gastric cancer stem cell (PD–GC‐SC) spheroids using an improved method. (A) Extracellular matrix (ECM) and Wnt ligand preference in primary culture. The spheroid lines were considered Wnt‐dependent when they perished in the cancer medium without L‐WRN CM during three serial passages. The three spheroid lines labeled with asterisks derived from a single patient. (B) Rapid establishment of GC‐SC spheroids in the improved culture condition. The duration time needed for expansion of each spheroid line from the patient sample to 12 wells of a 12‐well cell‐culture plate is plotted with the medians and interquartile ranges. p value, analyzed using Mann–Whitney U‐test. (C) Clinicopathological characteristics and mutational statuses of PD–GC‐SC spheroids. Shown are pathological features of 47 lines and representative genetic alterations of 43 lines. The pathological stage was determined by examination of surgically resected specimens. The HER2 status of the primary tumor was determined by immunohistochemistry or in situ hybridization. Cancer‐specific mutations were detected using a comprehensive cancer panel (HG1T–HG18T) or RNA sequencing (HG19T–HG47T). Indel, insertion/deletion variant; SNV, single nucleotide variant. The three spheroid lines labeled with asterisks derived from a single patient.