The role of extracellular conditions during CaCo-2 cells growth: a preliminary study for numerical model validation

Objectives: One important limitation in cell therapy protocols, and regenera- tive medicine (an innovative and promising strate- gy for different pathologies treatment), is the lack of knowledge about cells engraftment, prolifera- tion and differentiation. In order to allow an effi- cient and successful cell transplant, it is neces- sary to predict the logistics, economic and timing issues during cellular injection. It has been report- ed that several parameters, such as cells number, temperature and extracellular pH (pH0) value can influence metabolic pathways and cellular growth. Numerical analysis and model can help to reduce and understand the effects of the above environ- mental conditions on cell survival. The aim of this paper is to develop the first step of cells transplantation in order to identify "in vit- ro", which parameters can be useful to develop and validate a numerical model, able to evaluate "in vivo" cells engraftment and proliferation. Material and Methods: We studied the vari- ation of extracellular parameters - such as medi- um volume, buffer system, nutrient concentra- tions and temperature on human colon carcino- ma cells (CaCo-2) "in vitro culture" - pursuing the goal of understanding in deeper details cel- lular processes such as growth, metabolic activ- ity, survival and pH0. Results: Results showed that CaCo-2 cells growth and mortality increase after two days in culture when cells were suspended in 3.5 ml vol- ume to respect of 10 ml volume. Different tem- perature values influenced CaCo-2 cells growth and metabolic activity showing a direct relation- ship with the volume of the medium. Conclusions: Our results describe as CaCo-2 cell growth, metabolic activity, mortality and ex- tracellular pH were influenced by extracellular