Int. J. Dev. Biol. 62: 613 - 621 (2018)

https://doi.org/10.1387/ijdb.180161mf

Vol 62, Issue 9-10

A morphology-based assay platform for neuroepithelial-like cells differentiated from human pluripotent stem cells

Open Access | Original Article | Published: 10 October 2018

Mika Suga¹, Hiroaki Kii², Naoko Ueda¹, Yu-Jung Liu¹, Takako Nakano¹, Tomoro Dan², Takayuki Uozumi², Yasujiro Kiyota² and Miho K. Furue*^,1,2

¹Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka and ²Nikon Corporation, Tokyo, Japan

Abstract

Cell morphology is recognized as an important hallmark of neural cells. During the differentiation of human pluripotent stem cells (hPSCs) into neural cells, cell morphology changes dynamically. Therefore, characterization of the morphology of cells during this period is important to improve our understanding of the differentiation and development of neural cells. General methods for the directed induction of hPSCs include the steps of multi-cellular aggregation or high-density cell culture, particularly at the early phase of neural differentiation, and therefore, the morphology of each differentiating cell is difficult to recognize. Here, we have developed a new method for the directed differentiation of neuroepithelial-like cells (NELCs) from hPSCs at a low cell density in an adherent monolayer culture, as well as an image-processing algorithm to evaluate the cell morphology of differentiating NELCs, in order to follow cell morphology during the differentiation of hPSCs into NELCs. Using these methods, the morphological transition of differentiating cells was observed in real time using phase contrast imaging and then quantified. Because cell morphology is also considered an inherent biological marker of neural cells cultured in vitro, this method is potentially useful to study the mechanisms underlying neural cell differentiation.

Keywords

differentiation, neuroepithelial-like cell, monolayer culture, albumin-free defined culture medium