Gravesano, November 2020

Principles of in vitro nutrition for human stem cells

In the cultivation of stem cells in vitro, components of animal origin and/or chemically undefined extracts like human platelet lysate are still used to formulate culture media because the quality of the best known basal media is not adequate for cell culture in chemically defined environments. Experimental evidence shows that the better the qualitative and quantitative composition of the basal medium, the fewer additives of animal origin or chemically undefined components are needed to perform stem cell culture under chemically defined conditions.

The present comment deals with essential aspects concerning identity and quality of culture media components for pre-clinical and clinical research. It outlines the development of physico-chemically defined micro-environments for cell cultures that modulate functions of the metabolism of human mesenchymal stromal cells.

The development process typically consists of three parts. The first stage involves the development of fully defined micro-environments capable of satisfying the essential physiological requirements of the cell. For this purpose, a detailed analysis of the physical (pH, osmotic pressure) and chemical requirements (energy sources, amino acids, vitamins, precursors of fatty and nucleic acids, inorganic salts, trace elements and chemical impurities) of the cells is carried out. The second phase involves the total elimination of complex, chemically undefined variables such as fetal calf serum or human platelet lysate, which are to be considered as critical components in cell culture media. The selection of well balanced basal media and their manufacturing technique serve the purpose. The third and final stage involves the verification of the results by cultivation in bioreactors. These allow cell proliferation at significantly higher densities and extended control of the culture conditions by on-line analysis systems for the most important physical culture parameters (temperature, pH, partial oxygen pressure).

Lines of evidence indicate that good knowledge of the variables that influence cell growth processes has numerous advantages. The three-step strategy described above allows the identification of the essential physiological needs of the cells, thus favouring control and modulation of their main metabolic functions. This perspective becomes even more interesting as the omission of complex and expensive medium components in clinical culture processes leads to a reduction of contamination risks, product variability and manufacturing costs.

Ferruccio Messi, Ph. D
President and founder of Cell Culture Technologies

The above contribution was presented at the ISCT Paris Virtual Meeting, May 28-29, 2020.
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