Towards Physiologic Culture Approaches to Improve Standard Cultivation of Mesenchymal Stem Cells

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.

In Vitro Cellular and Molecular Interplay between Human Foreskin-Derived Mesenchymal Stromal/Stem Cells and the Th17 Cell Pathway

“…Furthermore, the preservation of the physiological characteristics of MSCs during their in vitro culture is essential for improving the efficiency of therapeutic and in vitro modeling applications, as recently reviewed [ 39 ]. In view of this document, there is considerable research focus on the optimization of specific culture media, culture conditions, and protocols for MSCs.…”

Processing and Ex Vivo Expansion of Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells for the Development of an Advanced Therapy Medicinal Product for use in Humans

“…During the selection of manufacturer of PL, it is necessary to verify the quality specification of the PL according to the aspects presented by Oeller et al [41]. Although on the market there are a few chemically defined media [42] and some of them were tested during our optimization research, we did not select one of them for use in our manufacturing process to simplify work (minimize the amount of manipulation it was necessary to conduct) in the cleanroom and to slightly reduce the manufacturing costs. To ensure the high quality of AT-MSCs, as the active substance of HE-ATMP, we conducted numerous cell seeding density assays to design a timeline of the manufacturing process.…”

Co-administration of human MSC overexpressing HIF-1α increases human CD34+ cell engraftment in vivo

“…However, maintaining in vitro the hypoxic conditions that MSC have physiologically in the human bone marrow niche is difficult to preserve only by regulating culture conditions, since the exposure to a normoxic environment rapidly reverses the gained features induced by hypoxia. There is extensive information about the key role of HIF-1α regulating the hypoxic metabolism [ 35 , 37 , 39 , 40 ]. That is why in the current work, we have opted by overexpressing HIF-1α in MSC by lentiviral transduction in order to mimic some of the effects induced by hypoxia and maintain those effects over time.…”