Page 14 - Shimadzu C2MAP System

Application Data

Optimization of Culture Processes and Scaling Up of Culture Volumes Cell Culture Profiling System

The combination of a High-Performance Triple Quadrupole Liquid Chromatograph Mass Spectrometer LCMS-8060/8050/8040
A culture supernatant after
and the Cell Culture Profiling Method Package enables multi-component, simultaneous analysis of the components of the
replacement of the culture media for
culture supernatant following manual pretreatment.
undifferentiated human iPS cells was
sampled. The temporal changes in the
components in the culture supernatant LC/MS/MS Method Package for Cell Culture Profiling
were then monitored using the C2MAP LCMS/8060/8050/8040
system. The results suggested that A 95-component simultaneous analysis, including amino
hypoxanthine and some other acids contained in culture media components and secreted
components were depleted from Day 2 metabolites, as well as sugars, vitamins, and organic acids,
or later, despite replacing the culture can be performed in 17 minutes per sample.
media on a daily basis. In addition, the
results indicated that asparagine,
pantothenic acid, folic acid and
pyridoxine maintained basically the LC/MS/MS Method Package for Primary Metabolites Ver. 2
same signal intensity throughout the Either the ion pair method (55 components), which targets important compounds
culture period, suggesting that they from the main metabolic pathways for biological samples, or the non-ion pair
are not easily consumed by the cells. method (97 components), which targets the main amino acids and organic acids,
Through multicomponent monitoring can be selected to suit the instrument environment.
of the culture supernatant
components, information can be
obtained regarding which components
are favored and consumed by cells, and LC/MS/MS MRM Library for Phospholipids Profiling
which are depleted during the culture
period. This information provides Analysis targets the main phospholipids in biological samples. Phospholipid
useful insights for optimizing the profiling is performed by combining two analysis methods: the phospholipid class
culture media composition and the determining method (422 components) and the fatty acid composition
culture process. determining method (867 components).

Next, the C2MAP system was used to compare the temporal changes in the culture supernatant components in LC/MS/MS Method Package for Lipid Mediators Ver. 2
undifferentiated human iPS cells and a model of cells deviated from the undifferentiated state (cytokines were added under
This can perform simultaneous analysis of 158 components, including the main
undifferentiated culture conditions to induce differentiation of the various germ layers). As a result, it was possible to find
lipid mediators, such as eicosanoids, polyunsaturated fatty acid metabolites, and
compounds indicating the characteristic temporal changes in each model. Such compounds can become marker candidates for
platelet activating factor.
use in performing culture process management.

Traverse MS ™

This software is for the high-speed analysis of MRM data from multiple samples
and multiple components. It can be used for principal component analysis and
hierarchical clustering. This software is available from Reifycs Inc.

Undifferentiation Endoderm Mesoderm Ectoderm
C2MAP System
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Cell Culture Media Analysis Platform