The C2MAP™ system measures component changes in a culture supernatant as culturing progresses using LC/MS/MS. This system can be used for a wide range of applications, from basic research of cell cultures including pluripotent stem cells (iPS cells and ES cells), mesenchymal stem cells, and antibody-producing cells, to scaling up of culture volumes and actual process development.

◆ Obtain Temporal Change Profiles for up to 95 Culture Supernatant Components

C2MAP system can provide simultaneous analysis methods for up to 95 components, making it suitable for component analysis to determine cell growth and status quality. The dedicated C2MAP TRENDS viewer software can graph component variations across multiple conditions for easier comparative analysis.

◆ Optimization of Culture Conditions

The system can provide useful insights for the optimization of culture conditions in animal cell cultures by monitoring the consumption and depletion of media components during culturing, as well as the variation in metabolites secreted from cells.

Features of the C2MAP System
1. Automated Process from Pretreatment to Measurement for the Culture Supernatant Analysis

With an automated process from pretreatment to measurement, automatic analysis can be performed at night and on non-working days.
The measurement workflow can be selected to match the actual culture.
Seamless analysis and management from pretreatment to LC/MS/MS measurement can be performed.
Pretreated samples are stocked on a microplate automatically to enable re-measurement with ease.
2. Supports a Wide Range of Measurement Compounds and Culture Supernatant Samples

A total of 95 components, including major basal culture media components for animal cells and secreted metabolites, can be simultaneously analyzed at high speed.
Applicable to a wide range of cell culture media
(iPS cells, ES cells, mesenchymal stem cells, T cells, and CHO cells)
3. Visualization of Component Variations in Culture Media

Temporal changes in the components can be displayed as trend graphs.
The results under multiple experimental conditions can be overlaid in the display, enabling comparative analysis.

For Research Use Only. Not for use in diagnostic procedures.

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Automated Pretreatment Module for Cell Culture Media Analysis

The Optimal Workflow Can Be Selected
The C2MAP system supports two modes: Batch mode (ordinarily used) that gives priority to the pretreatment of the culture supernatant samples placed, and sequential mode in which pretreatment and analysis are performed alternately for each sample.

Supports a Wide Range of Measurement Compounds and Culture Supernatant Samples
The analysis method stored in the control software allows the simultaneous analysis of the following 95 components plus 2-Isopropylmalic acid.

C2MAP TRENDS
For results obtained via LC/MS/MS, temporal changes in each component can be graphed by the dedicated viewer software. Analysts can monitor variations in metabolites secreted from cells and culture media components during cultivation, as well as display graphs of component comparisons with samples from different culture series. As a result, the consumption and depletion of culture media components, and changes in the amounts of metabolic components secreted from cells, can be observed, thereby providing useful insights into considerations of the optimal culture conditions and assessments of cellular status.

◆Temporal Changes in Measured Components

◆Measured-component Comparisons among Different Culture Series

For Research Use Only. Not for use in diagnostic procedures.

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◆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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For Research Use Only. Not for use in diagnostic procedures.

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