College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China.
Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Gansu Agricultural University, Lanzhou 730070, China.
Int J Mol Sci. 2023 Jan 18;24(3):1917. doi: 10.3390/ijms24031917.
Induced pluripotent stem cells (iPSCs) can differentiate into all types of cells and can be used in livestock for research on biological development, genetic breeding, and in vitro genetic resource conservation. The Bactrian camel is a large domestic animal that inhabits extreme environments and holds value in the treatment of various diseases and the development of the local economy. Therefore, we transferred four mouse genes (, , , and c-) into Bactrian camel fetal fibroblasts (BCFFs) using retroviruses with a large host range to obtain Bactrian camel induced pluripotent stem cells (bciPSCs). They were comprehensively identified based on cell morphology, pluripotency gene and marker expression, chromosome number, transcriptome sequencing, and differentiation potential. The results showed the pluripotency of bciPSCs. However, unlike stem cells of other species, late formation of stem cell clones was observed; moreover, the immunofluorescence of SSEA1, SSEA3, and SSEA4 were positive, and teratoma formation took four months. These findings may be related to the extremely long gestation period and species specificity of Bactrian camels. By mining RNA sequence data, 85 potential unique pluripotent genes of Bactrian camels were predicted, which could be used as candidate genes for the production of bciPSC in the future. Among them, , , , , , , , and are more attractive. In conclusion, we generated bciPSCs for the first time and obtained their transcriptome information, expanding the iPSC genetic information database and exploring the applicability of iPSCs in livestock. Our results can provide an experimental basis for Bactrian camel ESC establishment, developmental research, and genetic resource conservation.
诱导多能干细胞(iPSCs)可以分化为所有类型的细胞,并可用于家畜研究生物发育、遗传育种和体外遗传资源保存。双峰驼是一种大型家畜,栖息在极端环境中,在治疗各种疾病和发展当地经济方面具有价值。因此,我们使用具有广泛宿主范围的逆转录病毒将四个小鼠基因(,,, 和 c-)转入双峰驼胎儿成纤维细胞(BCFFs)中,获得了双峰驼诱导多能干细胞(bciPSCs)。我们基于细胞形态、多能性基因和标记物表达、染色体数目、转录组测序和分化潜能对它们进行了综合鉴定。结果表明 bciPSCs 具有多能性。然而,与其他物种的干细胞不同,观察到晚期形成干细胞克隆;此外,SSEA1、SSEA3 和 SSEA4 的免疫荧光呈阳性,并且畸胎瘤形成需要四个月。这些发现可能与双峰驼极长的妊娠期和物种特异性有关。通过挖掘 RNA 序列数据,预测了 85 个双峰驼潜在的独特多能基因,这些基因可作为未来生产 bciPSC 的候选基因。其中,,,,,,,, 和 更具吸引力。总之,我们首次生成了 bciPSCs 并获得了它们的转录组信息,扩展了 iPSC 遗传信息数据库,并探索了 iPSCs 在家畜中的适用性。我们的结果可为双峰驼 ESC 建立、发育研究和遗传资源保存提供实验基础。