Kim, Choong Lee, Kang Sun Bang, Jae Hoon Kim, Young Eyn Kim, Min-Cheol Oh, Kwang Wook Lee, Soo Hyun Kang, Ji Yoon 2024-01-20T17:32:01Z 2024-01-20T17:32:01Z 2021-09-04 2011-03 1473-0197 https://pubs.kist.re.kr/handle/201004/130572 This paper proposes a microfluidic device for the on-chip differentiation of an embryoid body (EB) formed in a microwell via 3-dimensional cultures of mouse embryonic carcinoma (EC) cells. The device adjusted the size of the EB by fluid volume, differentiated the EB by chemical treatment, and evaluated its effects in EC cells by on-chip immunostaining. A microfluidic resistance network was designed to control the size of the embryoid body. The duration time and flow rate into each microwell regulated the initial number of trapped cells in order to adjust the size of the EB. The docked cells were aggregated and formed a spherical EB on the non-adherent surface of the culture chip for 3 days. The EC cells in the EB were then differentiated into diverse cell lineages without attachment for an additional 4 days; meanwhile, retinoic acid (RA) was applied without serum to direct the cells into early neuronal lineage. On-chip immunostaining of the EB in the microwell with a neuronal marker was conducted to assess the differentiation-inducing ability of RA. The effect of RA on neuronal differentiation was analyzed with confocal microscopic images of the TuJ1 marker. The RA-treated cells expressed more neuronal markers and appeared as mature neuronal cells with long neurites. The fluorescence intensity of the TuJ1 in the RA-treated EB was twice that observed in the non-treated EB on day 5. It was demonstrated that the pre-screening of inducing chemicals on the early neuronal differentiation of EC cells in a single microfluidic chip was indeed feasible. This chip is expected to constitute a useful tool for assessing the early differentiation of ES cells without attachment, and is also expected to prove useful as an anti-cancer drug test platform for the cytotoxicity assay with cellular spheroids. English Royal Society of Chemistry IN-VITRO BODIES SIZE 3-Dimensional cell culture for on-chip differentiation of stem cells in embryoid body Article 10.1039/c0lc00516a 1 Lab on a Chip, v.11, no.5, pp.874 - 882 Lab on a Chip 11 5 874 882 scie scopus 000287409600014 2-s2.0-79952256268 Biochemical Research Methods Chemistry, Multidisciplinary Chemistry, Analytical Nanoscience & Nanotechnology Instruments & Instrumentation Biochemistry & Molecular Biology Chemistry Science & Technology - Other Topics Instruments & Instrumentation Article IN-VITRO BODIES SIZE cell culture stem cells embryoid body