Tada Masako, Morizane Asuka, Kimura Hironobu, Kawasaki Hiroshi, Ainscough Justin F X, Sasai Yoshiki, Nakatsuji Norio, Tada Takashi
Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Shogoin, Sakyo-ku, Kyoto, Japan.
Dev Dyn. 2003 Aug;227(4):504-10. doi: 10.1002/dvdy.10337.
Somatic nuclei can be epigenetically reprogrammed by factors present in undifferentiated embryonic stem (ES) cells. The acquisition of pluripotency by somatic genomes could render such cells a viable source of personalized cell type(s) for therapeutic application, avoiding the need for controversial therapeutic cloning. To investigate this possibility, we first determined the origin of transcripts in teratomas generated from mouse (ES x somatic cell) hybrid clones. Transcription of markers from the somatic genome demonstrated efficient in vivo differentiation down independent lineages. The induction of dopaminergic neurons by coculture with stromal PA6 feeder cells also demonstrated efficient capacity to differentiate in vitro. Hybrid clone-derived neurons expressed appropriate markers, and transcription of Pitx3 from the somatic genome was confirmed. When transplanted into mouse brains, the dopaminergic neurons were successfully integrated and expressed tyrosine hydroxylase. Thus, it should be possible to produce personalized ES-like cells with the reprogrammed somatic genomes.
体细胞的细胞核可以被未分化的胚胎干细胞(ES细胞)中存在的因子进行表观遗传重编程。体细胞基因组获得多能性可能使这些细胞成为用于治疗应用的个性化细胞类型的可行来源,从而避免了有争议的治疗性克隆的需要。为了研究这种可能性,我们首先确定了从小鼠(ES×体细胞)杂交克隆产生的畸胎瘤中转录本的来源。来自体细胞基因组的标志物的转录表明在体内能有效地沿着独立谱系分化。与基质PA6饲养细胞共培养诱导多巴胺能神经元也证明了其在体外有效的分化能力。杂交克隆来源的神经元表达了合适的标志物,并证实了来自体细胞基因组的Pitx3的转录。当移植到小鼠大脑中时,多巴胺能神经元成功整合并表达酪氨酸羟化酶。因此,利用重编程的体细胞基因组生产个性化的类ES细胞应该是可行的。