Stem Cell Biology and Regenerative Medicine Research Group, Research Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Stem Cell and Brain Research Institute, University of Lyon, Université Lyon 1, INSERM, INRAE, U1208, USC1361, 69500, Bron, France.
Sci Rep. 2023 Jul 29;13(1):12323. doi: 10.1038/s41598-023-39536-1.
The cultivation and expansion of chicken primordial germ cells (cPGCs) are of critical importance for both biotechnological applications and the management of poultry genetic biodiversity. The feeder-free culture system has become the most popular approach for the cultivation and expansion of cPGCs. However, despite some success in the cultivation of cPGCs, the reproducibility of culture conditions across different laboratories remains a challenge. This study aimed to compare two defined and enriched media for the growth of cPGCs originating from the Hubbard JA57 broiler. To this end, cPGCs were isolated from the embryonic blood of Hamburger-Hamilton (HH) stages 14-16 and cultured at various time points. The Growth properties and characteristics of these cells were evaluated in two different culture conditions (the defined or enriched medium) and their migratory properties were assessed after genetic engineering and injection into the vasculature of 2.5-day-old chicken embryos. The main finding of this study was that the use of an enriched medium (the defined medium with Knock-Out Serum Replacement; KOSR) resulted in improved growth properties of cPGCs originating from the Hubbard JA57 broiler compared to a defined medium. The ability to cultivate and expand cPGCs is crucial for the generation of both genetically engineered birds and breeds of interest from local or commercial origins. Therefore, these results highlight the importance of choosing an appropriate culture medium for cPGCs growth and expansion.
鸡原始生殖细胞(cPGCs)的培养和扩增对于生物技术应用和家禽遗传生物多样性的管理都至关重要。无饲养层培养系统已成为培养和扩增 cPGCs 的最流行方法。然而,尽管在 cPGCs 的培养方面取得了一些成功,但不同实验室之间培养条件的可重复性仍然是一个挑战。本研究旨在比较两种用于生长源自 Hubbard JA57 肉鸡的 cPGCs 的定义和丰富培养基。为此,从 Hamburger-Hamilton(HH)阶段 14-16 的胚胎血液中分离出 cPGCs,并在不同时间点进行培养。在两种不同的培养条件(定义或丰富培养基)下评估这些细胞的生长特性和特征,并在遗传工程后评估其迁移特性和注射到 2.5 天大的鸡胚血管中。本研究的主要发现是,与定义培养基(含 Knock-Out Serum Replacement 的定义培养基;KOSR)相比,使用丰富培养基可改善源自 Hubbard JA57 肉鸡的 cPGCs 的生长特性。培养和扩增 cPGCs 的能力对于生成遗传工程鸟类和来自本地或商业来源的感兴趣品种都至关重要。因此,这些结果强调了选择合适的培养基对于 cPGCs 生长和扩增的重要性。