Microencapsulation is one of the promising strategies to develop a three-dimensional in vivo tumour-mimic model in cancer research. Although previous studies have shown that tumour cells grow well during the microencapsulated culture, it is still not clear whether the electrostatic encapsulation process has an important effect on cellular characteristics. In this study, we investigated cellular response against non-physiological stress factors existing in the electrostatic microencapsulation process, such as the high-voltage electrostatic field, suspension and nutrition-free status. Our results showed that these non-physiological stress factors did not significantly induce cellular apoptosis, and did not affect cellular adhesion and viability. Furthermore, no change was found about invasion and drug resistance of the tumour cells. The normal endoplasmic reticulum function might play a role in maintaining biological properties during the electrostatic microencapsulation process.