2-dimensional (2D) co-cultures show limited resemblence with in vivo microenvironment while 3-dimensional (3D) co-cultures form a micromass which is more similar with in vivo microenvironmentthus it would be more useful in biomedical research. This study was intended to compare 2D co-cultures with 3D co-cultures of stem cells derived from umbilical cord blood vessels and hepatic stellate cells, conducted with hanging drop method to assess the cell morphology and the formation of spheroid from the micromass.. This in vitrostudy was conducted at Institute of Human Virology and Cancer Pathobiology (IHVCB) UI and histology laboratory from September 2015 to October 2016 using stem cells which were isolated from human umbilical cord blood (UCB) and LX-2 cell line (human hepatic stelate cells). Human umbilical cord blood was sorted with MACS CD34 and percentage of CD34+ cells were analyzed by flowcytometry. Stem cell co-cultures (UCB) or umbilical cord and L2 was did by hanging drop methods for 2D co-culture and 3D co-culture. Triplicates experiments were performed for each set of co-culture. The results showed the difference in the morphology of 2D co-culture and 3D hanging drop compared to monoculture. In the 2D co-culture there was a micromass formation, whereas in the 2D monoculture, the micromass was not formed. In the 3D hanging drop there was a smaller spheroid compared to the 3D hanging drop monoculture. The morphology of 2D and 3D co-culture cells with hanging drop method in comparison with monoculture cells showed phenotypic changes of cells which were incorporated together in the micromass.