Visualization of morphological categories of colonies for monitoring of effect on induced pluripotent stem cell culture status

• Main Authors: Risako Nagasaka, Megumi Matsumoto, Mai Okada, Hiroto Sasaki, Kei Kanie, Hiroaki Kii, Takayuki Uozumi, Yasujiro Kiyota, Hiroyuki Honda, Ryuji Kato
• Format: Article
• Language: English
• Published: Elsevier 2017-06-01
• Series: Regenerative Therapy
• Subjects: Human induced pluripotent stem cells; Morphological analysis; Colony morphology; Culture technique; Colony morphological diversity
• Online Access: http://www.sciencedirect.com/science/article/pii/S235232041630058X

From the recent advances, there are growing expectations toward the mass production of induced pluripotent stem cells (iPSCs) for varieties of applications. For such type of industrial cell manufacturing, the technology which can stabilize the production efficiency is strongly required. Since the present iPSC culture is covered by delicate manual operations, there are still quality differences in produced cells from same culture protocols. To monitor the culture process of iPSCs with the quantified data to evaluate the culture status, we here introduce image-based visualization method of morphological diversity of iPSC colonies. We have set three types of experiments to evaluate the influential factors in iPSC culture technique that may disturb the undifferentiation status of iPSC colonies: (Exp. 1) technical differences in passage skills, (Exp. 2) technical differences in feeder cell preparation, and (Exp. 3) technical differences in maintenance skills (medium exchange frequency with the combination of manual removal of morphologically irregular colonies). By measuring the all existing colonies from real-time microscopic images, the heterogenous change of colony morphologies in the culture vessel was visualized. By such visualization with morphologically categorized Manhattan chart, the difference between technical skills could be compared for evaluating appropriate cell processing.