◆Optimization of Culture Processes and Scaling Up of Culture Volumes
A culture supernatant after replacement of the culture media for undifferentiated human iPS cells was sampled. The temporal changes in the components in the culture supernatant were then monitored using the C2MAP system. The results suggested that hypoxanthine and some other components were depleted from Day 2 or later, despite replacing the culture media on a daily basis. In addition, the results indicated that asparagine, pantothenic acid, folic acid and pyridoxine maintained basically the same signal intensity throughout the culture period, suggesting that they are not easily consumed by the cells. Through multicomponent monitoring of the culture supernatant components, information can be obtained regarding which components are favored and consumed by cells, and which are depleted during the culture period. This information provides useful insights into optimization of the culture media composition and the culture process.
Next, the C2MAP system was used to compare the temporal changes in the culture supernatant components in undifferentiated human iPS cells and a model of cells deviated from the undifferentiated state (cytokines were added under undifferentiated culture conditions to induce differentiation of the various germ layers). As a result, it was possible to find compounds indicating the characteristic temporal changes in each model. Such compounds can become marker candidates for use in performing culture process management.
◆Qualitative Analysis of Basal Culture Media / Culture Media Additives
It is known that a difference in culture media additives has an impact on the culture results. In the example below, various types of Dulbecco’s modified Eagle medium (DMEM) were analyzed by LC-MS/MS. The following table shows the differences in the DMEM culture media compositions noted in the product catalog.
The results of the analysis match the content noted in the product catalog. When culture media and other raw materials are received, confirming the existence of the components that they should contain is important for stable substance production. Furthermore, creating calibration curves using standard products makes it possible to also quantify the concentrations, so the C2MAP system can be applied to the quantitative analysis of culture media.
For Research Use Only. Not for use in diagnostic procedures.
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