This paper presents the formation of double-layer cellular tumor spheroid for the co-culture of hepatocyte (HepG2 cancer liver cell and LO2 normal liver cell) using alginate hydrogel. With the microfluidic chip, double-layer cellular tumor spheroid can be formed by the hydrogel droplet to mimic the in vivo environment in which the cancer cell is surrounded by normal cell. A microchip with concentration gradient generation and tumor spheroid trapping site is designed for anti-cancer drug analysis.