Cha H-J, Yun J I, Han N R, Kim H-Y, Baek S, Lee S-H, Lee J, Lee E, Park C-K, Lee S T
Department of Animal Life Science, Kangwon National University, Chuncheon, Korea.
College of Veterinary Medicine, Institute of Veterinary Science, Kangwon National University, Chuncheon, Korea.
Reprod Domest Anim. 2018 Feb;53(1):176-185. doi: 10.1111/rda.13088. Epub 2017 Nov 6.
Although basic fibroblast growth factor (bFGF) is an essential factor supporting the maintenance of porcine embryonic stem (ES) cell self-renewal and pluripotency, its high cost has limited previous studies, and the development of a low-cost culture system is required. For these systems, in vivo blastocysts were progressively cultured under various conditions consisting of different culture mediums and/or different feeder cell numbers at a low concentration of bFGF. As the results, the sequential culture of in vivo-derived porcine blastocysts on 5.0 × 10 mouse embryonic fibroblast (MEF) feeder cells in alpha minimum essential medium-based medium for primary culture, on 2.5 × 10 MEF feeder cells in Mixture medium for the 1st subpassage, and on 2.5 × 10 MEF feeder cells in DMEM/Ham's F10-based medium for the post-2nd subpassage could support the establishment and maintenance of porcine ES-like cells at the low concentration of bFGF. The established porcine ES-like cells showed ES cell-specific characteristics such as self-renewal and pluripotency. We confirmed that porcine ES-like cells could be generated from in vivo-derived porcine blastocysts at a low concentration of bFGF.
尽管碱性成纤维细胞生长因子（bFGF）是维持猪胚胎干细胞（ES）自我更新和多能性的关键因子，但其高昂成本限制了以往的研究，因此需要开发低成本培养系统。对于这些系统，将体内囊胚在低浓度bFGF条件下，于由不同培养基和/或不同数量饲养细胞组成的各种条件下进行逐步培养。结果表明，将体内来源的猪囊胚先后培养在以α-最低必需培养基为基础的培养基中、5.0×10个小鼠胚胎成纤维细胞（MEF）饲养细胞上进行原代培养，在混合培养基中、2.5×10个MEF饲养细胞上进行第1次传代培养，以及在以DMEM/Ham's F10为基础的培养基中、2.5×10个MEF饲养细胞上进行第2次传代后培养，能够在低浓度bFGF条件下支持猪ES样细胞的建立和维持。所建立的猪ES样细胞表现出ES细胞特异性特征，如自我更新和多能性。我们证实，低浓度bFGF条件下可从体内来源的猪囊胚生成猪ES样细胞。
