Abstract
Mesenchymal stem cells (MSCs) are of great interest for their use in cell-based therapies due to their multipotent differentiation and immunomodulatory capacities. In consequence of limited numbers following their isolation from the donor tissue, MSCs require extensive expansion performed in traditional 2D cell culture setups to reach adequate amounts for therapeutic use. However, prolonged culture of MSCs in vitro has been shown to decrease their differentiation potential and alter their immunomodulatory properties. For that reason, preservation of these physiological characteristics of MSCs throughout their in vitro culture is essential for improving the efficiency of therapeutic and in vitro modeling applications. With this objective in mind, many studies already investigated certain parameters for enhancing current standard MSC culture protocols with regard to the effects of specific culture media components or culture conditions. Although there is a lot of diversity in the final therapeutic uses of the cells, the primary stage of standard isolation and expansion is imperative. Therefore, we want to review on approaches for optimizing standard MSC culture protocols during this essential primary step of in vitro expansion. The reviewed studies investigate and suggest improvements focused on culture media components (amino acids, ascorbic acid, glucose level, growth factors, lipids, platelet lysate, trace elements, serum, and xenogeneic components) as well as culture conditions and processes (hypoxia, cell seeding, and dissociation during passaging), in order to preserve the MSC phenotype and functionality during the primary phase of in vitro culture.
Highlights
Cell-based therapies aim to repair or regenerate defective tissues or organs due to physical or congenital damage, ageing, and degenerative diseases
In consequence of limited numbers following their isolation from the donor tissue, Mesenchymal stem cells (MSCs) require extensive expansion performed in traditional 2D cell culture setups to reach adequate amounts for therapeutic use
In contrast with these results, a few studies have reported that high glucose does not have an effect on the apoptosis and proliferation rates of adiposederived [52] and primary bone marrow MSCs (BM-MSCs) [53], while it can significantly enhance proliferation of telomerase-immortalized MSCs
Summary
Cell-based therapies aim to repair or regenerate defective tissues or organs due to physical or congenital damage, ageing, and degenerative diseases. For that reason, during the last years, concerns over the biological relevance and reproducibility of in vitro culture techniques have led to the development of advanced three-dimensional (3D) and dynamic cell culture methods. These methods serve as better models for the representation of in vivo physiological conditions [13,14], as well as platforms for upscaling production of cells for clinical applications [15,16]. We want to review on optimizing approaches for standard 2D in vitro expansion of human MSCs, focusing on culture media components (amino acids, ascorbic acid, glucose level, growth factors, lipids and fatty acids, platelet lysate, trace elements, serum, Cells 2021, 10, 886 and xenogeneic components) as well as culture conditions and processes (hypoxia, seeding density and cell dissociation during passaging)
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