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April 15, 2010 (Vol. 30, No. 8)

Automated Stem Cell Culture with a Twist

Researchers Develop and Refine a System for Higher Consistency and Reproducibility

  • The Automation Solution

    Click Image To Enlarge +
    Figure 2. Variations in temperature and pH of liquid culture medium as microplates are manually transported between a CO2 incubator at 37ºC and a biological containment cabinet with no environmental control.

    Automation provides a comprehensive solution to inconsistencies in manual handling, ensuring that every step of the process is performed in the same way, for the same duration, every time. In addition to improving the consistency of operations, it is possible to perform the entire culture procedure within a precisely controlled environment, avoiding fluctuation in temperature and pH.

    The team at UCL has developed a fully automated stem cell cultivation platform based on a Freedom EVO® liquid-handling workstation (Tecan). This integrated system is enclosed within a specially developed Class 2 biological containment cabinet—allowing complete environmental control—and incorporates all the equipment required for fully automated stem cell culture and manipulation, including pipetting and robotic manipulator arms, a microplate centrifuge, and a CO2 incubator.

    The sterile, temperature-controlled environment within the cabinet allows independent regulation of atmospheric gases during culture, particularly O2 and CO2. Conditions inside the cabinet are maintained at 37ºC, 5% CO2, and 21% O2, with no variations in temperature, CO2, or pH as plates are moved from the incubator to the deck of the instrument for processing.

    This minimizes fluctuations in environmental conditions that have been shown to impact on both cell growth over multiple passages and on the ultimate differentiation into specific cell types. The entire culture operation is performed using disposable pipette tips and consumables to minimize the risks of carryover and contamination and maximize operator safety when dealing with patient-derived cells.

  • Consistency and Reproducibility

    Click Image To Enlarge +
    Figure 3. Automated culture of pluripotent stem cells over eight consecutive passages

    Using the Tecan platform, the UCL team has shown that automated processing improves the consistency and reproducibility of pluripotent stem cell culture compared to manual processes performed by a highly skilled researcher.

    Figure 3 shows the automated culture of pluripotent stem cell populations through several passages. While both techniques gave similar cell numbers, automation improved homogeneity of the stem cell population, with no significant changes in gene-expression profile, cell viability, or cell density between successive passages.

  • Conclusions

    Automation offers reliable and consistent production of pluripotent stem cells and differentiated cell types, allowing the controlled and reproducible study of stem cell biology. It also provides a solution for industrial scale-out production, contributing to the commercial development of stem cell technologies for drug discovery and cell-based therapies.

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