Nano Medical Engineering Laboratory, RIKEN Cluster for Pioneering Research, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan; Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
Bioengineering Laboratory, RIKEN Cluster for Pioneering Research, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
Biochem Biophys Res Commun. 2019 Dec 3;520(2):257-262. doi: 10.1016/j.bbrc.2019.09.131. Epub 2019 Oct 5.
Based on a previous finding that fusion of a somatic cell with an embryonic stem (ES) cell reprogrammed the somatic cell, genes for reprogramming transcription factors were selected and induced pluripotent stem (iPS) cell technology was developed. The cell fusion itself produced a tetraploid cell. To avoid nuclear fusion, a method for cytoplasmic fusion using a microtunnel device was developed. However, the ES cell was too small for cell pairing at the device. Therefore, in the present study, ES cell enlargement was carried out with the colchicine derivative demecolcine (DC). DC induced enlargement of ES cells without loss of their stemness. When an enlarged ES cell was paired with a somatic cell in the microtunnel device, cytoplasmic fusion was observed. The present method may be useful for further development of reprogramming techniques for iPS cell preparation without gene transfection.
基于之前的一项发现,即体细胞与胚胎干细胞(ES 细胞)融合可重新编程体细胞,因此选择了重编程转录因子的基因,并开发了诱导多能干细胞(iPS 细胞)技术。细胞融合本身产生了四倍体细胞。为了避免核融合,开发了一种使用微隧道装置的细胞质融合方法。然而,ES 细胞太小,无法在设备中配对。因此,在本研究中,使用秋水仙碱衍生物秋水仙素(DC)对 ES 细胞进行了扩增。DC 诱导 ES 细胞增大而不丧失其干性。当增大的 ES 细胞与微隧道装置中的体细胞配对时,观察到细胞质融合。本方法可能有助于进一步开发无需基因转染的 iPS 细胞制备重编程技术。