Saito Ayaka, Kato Hidemasa, Kiyosawa Hidenori
Laboratory for Genomic Sciences, Department of Life Science, Chiba Institute of Technology, 2-17-1 Tsudanuna, Narashino, Chiba 275-0016, Japan.
Department of Developmental Biology and Functional Genomics, Graduate School of Medicine, Ehime University, 454 Shitsukawa, Toon, Ehime 791-0295, Japan.
Exp Anim. 2025 Apr 20;74(2):143-150. doi: 10.1538/expanim.24-0094. Epub 2024 Oct 15.
Allele-specific, monoallelic expression in diploid organisms represents an extreme case of allelic imbalance resulting from incompatibility between cis- and trans-elements. Due to haploinsufficiency, such monoallelic expression can lead to sporadic genetic diseases. In mice, allelic imbalances can be introduced into F1 offspring from inbred strains. Previously, we established F1 hybrid embryonic stem (ES) cell lines derived from four different mouse strains, each belonging to a different subspecies with substantial genetic polymorphisms. In this study, we investigated the neural differentiation capacity of the established ES cell lines. By introducing different culture conditions, which kept the ES cells undifferentiated under various pluripotencies, we succeeded in differentiating the majority of ES cell lines (eight out of eleven) with our default neural differentiation paradigm. Still, three lines exhibited insufficient differentiation despite combining culture conditions promoting undifferentiated as well as differentiated status. In addition, Ube3a imprinting was seen in two lines. Our findings contribute to the methodological understanding of mouse ES cell pluripotency and lead to the practical utility of F1 hybrid ES cells as a model for studying phenotypes resulting from gene locus interactions.
在二倍体生物中,等位基因特异性的单等位基因表达代表了由顺式和反式元件之间不兼容导致的等位基因失衡的极端情况。由于单倍剂量不足,这种单等位基因表达可导致散发性遗传疾病。在小鼠中,等位基因失衡可引入自交系的F1后代中。此前,我们建立了源自四种不同小鼠品系的F1杂交胚胎干细胞(ES)系,每个品系属于具有大量遗传多态性的不同亚种。在本研究中,我们研究了所建立的ES细胞系的神经分化能力。通过引入不同的培养条件,这些条件在各种多能性状态下使ES细胞保持未分化状态,我们成功地利用我们默认的神经分化模式使大多数ES细胞系(11个中的8个)分化。然而,尽管结合了促进未分化和分化状态的培养条件,仍有三个细胞系表现出分化不足。此外,在两个细胞系中观察到Ube3a印记。我们的研究结果有助于从方法学上理解小鼠ES细胞的多能性,并使F1杂交ES细胞作为研究基因座相互作用导致的表型的模型具有实际应用价值。