Kurosawa Hiroshi, Imamura Tetsuya, Koike Mikiko, Sasaki Katsunori, Amano Yoshifumi
Life Environment Medical Engineering, Division of Medical and Engineering Science, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Takeda, Kofu 400-8511, Japan.
J Biosci Bioeng. 2003;96(4):409-11. doi: 10.1016/S1389-1723(03)90148-4.
We proposed a simple method for forming an embryoid body (EB) from mouse embryonic stem (ES) cells using a polypropylene 1.5-ml conical tube with a screw cap. An ES cell suspension containing 2 x 10(4) cells was incubated in a conical tube. After 5 d of incubation, a single EB of 440 microm average diameter was formed in the conical tube. The formation efficiency of EB, which is the ratio of the number of tubes showing EB formation to the number of tubes seeded with ES cells, was greater than 99% in the conical tube, while it was approximately 60% in a hanging drop culture. The 5-day-old EB formed by the conical tube method had a sufficient differentiation ability. The beating of the cardiac muscle was microscopically observed in the populations derived from the 5-day-old EB.
我们提出了一种使用带螺帽的1.5毫升聚丙烯锥形管从小鼠胚胎干细胞形成胚状体（EB）的简单方法。将含有2×10⁴个细胞的胚胎干细胞悬液在锥形管中培养。培养5天后，在锥形管中形成了平均直径为440微米的单个胚状体。胚状体的形成效率是指显示胚状体形成的管数与接种胚胎干细胞的管数之比，在锥形管中大于99%，而在悬滴培养中约为60%。通过锥形管法形成的5日龄胚状体具有足够的分化能力。在源自5日龄胚状体的细胞群体中通过显微镜观察到了心肌的跳动。
