Investigation of Growth Conditions for the Expansion of Porcine Mesenchymal Stem Cells on Microcarriers in Stirred Cultures

Abstract

The extensive use of mesenchymal stem cells (MCS) in tissue engineering and cell therapy increases the necessity to improve their expansion. Among these, porcine MCS are valuable models for tissue engineering and are classically expanded in static T-flasks. In this work, different processes of stirred cultures were evaluated and compared. First, the effect of glucose, glutamine, antioxidant, and growth factors concentrations on porcine MSC expansion were analyzed in a suitable medium by performing kinetic studies. Results showed that a lower glucose concentration (5.5 mM) enabled to increase maximal cell concentration by 40 % compared with a higher one (25 mM), while addition of 2 to 6 mM of glutamine increased maximal cell concentration by more than 25 % compared with no glutamine supplementation. Moreover, supplementation with 1 μM thioctic acid increased maximal cell concentration by 40 % compared with no supplementation. Using this adapted medium, microcarriers cultures were performed and compared with T-flasks expansion. Porcine MSC were shown to be able to proliferate on the five types of microcarriers tested. Moreover, cultures on Cytodex 1, Cytopore 2, and Cultispher G exhibited a MSC growth rate more than 40 % higher compared with expansion in T-flasks, while MSC metabolism was similar.

Growth of human mesenchymal stem cells (MSCs) on films of enzymatically modified chitosan

“…In fact, normal cells especially stem cells are highly sensitive to ROS, thus the adding of antioxidant agent in culture medium is useful to reduce ROS‐mediated damage in cell systems such as neuronal cell systems . Several studies showed that the presence of antioxidant compounds in the basal medium, such as ascorbic acid, thioctic acid, β‐mercaptoethanol and sodium selenium, has positive effects on MSCs growth …”

“…To avoid cell aggregation, some studies proposed controlling operating parameters such as agitation rate, cell density, and addition of carriers . Sometimes, due to the smooth surface of microcarriers such as glass microcarriers and Cytodex, MSCs cannot attach firmly and may detach and be washed away during medium changes . In addition, those microcarriers cannot be digested in the human body .…”

Mass Production of Mesenchymal Stem Cells — Impact of Bioreactor Design and Flow Conditions on Proliferation and Differentiation

“…The impact of glutamine has been studied less extensively than that of glucose. Ferrari et al observed an increase in peak cell density by approx 25 % when adding of 2 to 6 mM of glutamine [84]. Schop et al concluded from their studies that glutamine has no importance as an energy source for hMSCs [93].…”

“…Due to the importance of hyperglycemia in vivo and the hypothesis, that high glucose could be deleterious to stem cell therapy, the impact of glucose on proliferation, potential for differentiation, and other physiological effects have been studied intensively. Glucose concentration in basal media has been shown to have both positive and negative impacts on MSC growth [47,[82][83][84][85][86][87][88][89][90][91][92][93]. It is beyond the scope of this chapter to discuss the various effects in detail but certain findings are presented below to highlight the complexity of this topic:…”

“…• Increase in peak cell density by 40 % at low glucose concentration (5.5 mM) when compared with an expansion at high glucose concentration (25 mM) [84].…”