Culture medium for esophageal squamous cell carcinoma epithelial cells, culture method, and application thereof

A primary cell culture medium for culturing primary esophageal squamous cell carcinoma (ESCC) epithelial cells,

• comprising a MST1/2 kinase inhibitor;
• a ROCK inhibitor;
• epidermal growth factor;
• insulin-transferrin-selenium complex;
• hepatocyte growth factor;
• a TGFβ type I receptor inhibitor selected from at least one of A83-01, SB431542, Repsox, SB505124, SB525334, SD208, LY36494, and SJN2511;
• and a P38/MAPK inhibitor selected from at least one of SB202190, SB203580, VX-702, VX-745, PD169316, RO4402247, and BIRB796;
• wherein the MST1/2 kinase inhibitor comprises a compound of Formula (I) or a pharmaceutically acceptable salt, or a solvate thereof, and the ROCK inhibitor is at least one selected from the group consisting of Y27632, Fasudil, and H-1152,
• wherein,
• R1 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, C4-C8 cycloalkylalkyl, C2-C6 spirocycloalkyl, and aryl optionally independently substituted with 1-2 R6, aryl C1-C6 alkyl optionally independently substituted with 1-2 R6 and heteroaryl optionally independently substituted with 1-2 R6;
• R2 and R3 are each independently selected from C1-C6 alkyl;
• R4 and R5 are each independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, C4-C8 cycloalkylalkyl, hydroxyl C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkylamino C1-C6 alkyl, C1-C6 alkoxy C1-C6 alkyl, and C3-C6 heterocyclyl C1-C6 alkyl;
• and
• R6 is selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, and C1-C6 haloalkyl.