Evaluación de la expansión de células madre mesenquimales en condiciones hipóxicas y medio suplementado con plasma pobre en plaquetas para la posible aplicación en la medicina regenerativa

Abstract

Mesenchymal stem cells (MSC) are used in regenerative medicine to treat various diseases. However, during its expansion in vitro, a high percentage dies due to standard culture conditions or time. For culture, medium supplemented with fetal bovine serum is usually used, which can cause immune problems, contamination and which, in turn, is expensive. In addition, apart from the incubation at 37 °C, the environment is in normal atmospheric oxygen conditions (normoxia) that differs from the physiological niche of the cells (hypoxia). Despite the known limitations, these methods continue to be used without taking into account the collateral damage that may be caused to the cells. Therefore, in this project the effect of hypoxic conditions (5% O2) and non-xenogeneic medium such as platelet-poor plasma (PPP) was evaluated by analyzing the number of population generations and population doubling time during the study. time in culture, in order to determine if the conditions presented above are optimal or feasible in the implementation of cell cultures. The evaluation was performed using four different culture conditions, which comprise normoxia with FBS as control group, normoxia with PPP, hypoxia with FBS and hypoxia with PPP. Through this research, we sought to evaluate the effect of the proposed experimental conditions for the expansion of MSCs, establishing the time and rate of population doubling. The control presented a proliferation rate of 0.989 and the cultures with the experimental conditions had a rate greater than 0.940, the condition in hypoxia supplemented with FBS being closest to the control with a value of 0.978. In addition, considering the population doubling time (TDP) reached by the MSCs in each one of the conditions, it was obtained that in a time of 1057 hours the cells supplemented with PPP did not exceed 15 population generations (GP) while in the same time the control reached 23 GP and in hypoxia with FBS approximately 22 GP. From this it is concluded that the culture condition in hypoxia with FBS is the one that presents more similarities to the control and can improve cell expansion, reduce oxidative stress, among others. On the other hand, the use of PPP is not recommended since its doubling time is considerably higher than the control.

Date of Award	Jul 2023
Original language	Spanish (Colombia)
Awarding Institution	Universidad Autónoma de Bucaramanga Ingeniería Biomédica
Supervisor	Silvia Milena Becerra Bayona (Supervisor) & Víctor Alfonso Solarte David (Supervisor)