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January 26, 2024
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This study focuses on automating the culture of iPS cells. Our goal is to reduce the variability due to manual experimentations and human labor by utilizing smaller and more affordable equipment, which leads to enhanced reproducibility and provides opportunities for more researchers who are even unfamiliar with iPS cells. Several types of automated culture machines have emerged, but most of them are still large, costly, and can only perform partial tasks.
The simple structure of the working arm has reduced the size and cost of the equipment. In addition, the tasks from cell maintenance to differentiation induction can be performed automatically with only prior material preparation and task setting on software. Machine repetition is remarkably accurate compared to human repetition.
The use of such machines not only improves reproducibility but also reduce human labor. Furthermore, if the parameters in each protocol are shared, researchers unfamiliar with iPS cells will be able to achieve similar results as well-experienced researchers, making it possible for them to easily enter into research using iPS cells. We are exploring the possibility of incorporating an AI-based image judgment system to automate the selection of the next work schedule, setting date and work contents from the learning of the experiment results.
We also anticipate that allowing researchers to tailor their working protocols to their preferences and share them online will not only enhance reproducibility among the world researchers, but also encourage global research collaboration. To begin loading, switch on the automated cell culture instrument and launch the operating software. Click on the loading button on the software, followed by loading preparation start.
Now place the dishes into the apparatus in their specific positions. Close the front sliding window and push the mechanical safety confirmation button. Choose the type and quantity of dishes in the software.
Then click on loading preparation completed, followed by the loading start button. Once the dish has been uploaded into the system, register notes about each individual dish in the software. Then click on registration to complete the loading operation.
To unload the dishes, first click on unload. Then select the dishes to be removed. Next, press the unloading preparation start button and click on unloading start.
When the instrument transfers the dishes from the incubator to the workbench, click on the dish-removal button. Manually open the front sliding window and remove the dishes. Immediately shut the front sliding window and push the safety confirmation button to complete unloading.
To replenish consumables, click on the consumables button on the software. For the pipettes, click on the pipette button followed by the replenish button. Choose the rack to be replenished.
Then press replenish start. Manually open the sliding rack window and replenish the pipettes as needed. Then close the window and push the mechanical safety button.
Click on the replenish completed button followed by the replenishment setup to input the information. Then click on the registration button and the replenish-completed button to finish. Similarly, replenish the tubes and media as desired.
Then click on the close button to exit the replenishment setup. To select tasks, click on the task button on the software. Now press the task setting button and choose the desired task from the task list.
Then click on the next step button. Input the date and time of the task to be performed. Then click on the registration button.
Now select the dish or plate to perform the task. Then click on the registration button again. Reconfirm the selected task and press registration.
Then click on start to automatically begin the task at the specified time and date. To cancel a scheduled task in advance, click on stop. Then choose the task to be aborted.
Click on edit task followed by the task cancel button and confirm the task deletion. To perform cell passaging and differentiation, choose the desired culture settings as per the desired cell to be obtained. The automated cell culture allowed for smooth cell proliferation over time.
Immunocytometric analysis showed that the induced pluripotent stem cells remained undifferentiated. This was confirmed by immunohistochemical analyses. No allelic abnormalities were observed after five maintenance cultures.
The pluripotent stem cells were observed to undergo differentiation into cardiomyocytes, hepatocytes, neuronal precursor cells, and keratinocytes with the automated system.
Here, we present a protocol for an automated cell culture system. This automated culture system reduces labor and benefits the users, including researchers unfamiliar with handling induced pluripotent stem (iPS) cells, from the maintenance of iPS cells to differentiation into various types of cells.
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Cite this Article
Bando, K., Yamashita, H., Hattori, F. An Automated Culture System for Maintaining and Differentiating Human-Induced Pluripotent Stem Cells. J. Vis. Exp. (203), e65672, doi:10.3791/65672 (2024).
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