Canine induced pluripotent stem cell maintenance under feeder-free and chemically-defined conditions,Molecular Reproduction and Development

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Canine induced pluripotent stem cell maintenance under feeder-free and chemically-defined conditions
Molecular Reproduction and Development ( IF 2.7 ) Pub Date : 2021-05-19 , DOI: 10.1002/mrd.23478
Kazuto Kimura ₁ , Masaya Tsukamoto ₁ , Takumi Yoshida ₁ , Miyuu Tanaka ₂ , Mitsuru Kuwamura ₂ , Manami Ohtaka ₃ , Ken Nishimura ₄ , Mahito Nakanishi _{3,

5} , Kikuya Sugiura ₁ , Shingo Hatoya ₁

Affiliation

Canine induced pluripotent stem cells (ciPSCs) provide a platform for regenerative veterinary medicine, disease modeling, and drug discovery. However, in the conventional method, ciPSCs are maintained using chemically-undefined media containing unknown animal components under on-murine embryonic fibroblast feeder conditions, which were reported to modify cell surface of iPSCs and increases the risk of immune rejection when the cells are transplanted into patients. Moreover, in the conventional method, ciPSCs are mechanically passaged, which requires much time and effort. Therefore, the large-scale expansion of ciPSCs is difficult, which should be resolved for using ciPSCs in clinical application and research. Here, it was shown that StemFit® AK02N and iMatrix-511 could maintain the pluripotency of ciPSCs using conventional culture method. Furthermore, it was demonstrated that the feeder-free and chemically-defined ciPSC culture systems using StemFit® AK02N and iMatrix-511 could stably maintain and allow the easy expansion of ciPSCs generated using N2B27 and StemFit® AK02N, without causing karyotype abnormalities. ciPSCs expressed several pluripotency markers and formed teratomas, including cells derived from three germ layers.

中文翻译：

在无饲养层和化学定义条件下犬诱导的多能干细胞维持

犬诱导多能干细胞 (ciPSC) 为再生兽医学、疾病建模和药物发现提供了一个平台。然而，在传统方法中，在小鼠胚胎成纤维细胞饲养条件下，使用含有未知动物成分的化学不确定培养基来维持 ciPSCs，据报道，这会改变 iPSCs 的细胞表面并增加细胞移植时免疫排斥的风险。耐心。此外，在传统方法中，ciPSCs 是机械传代的，这需要很多时间和精力。因此，ciPSCs的大规模扩容困难，在临床应用和研究中使用ciPSCs应该解决这个问题。在这里，表明 StemFit® AK02N 和 iMatrix-511 可以使用传统培养方法维持 ciPSC 的多能性。此外，研究表明，使用 StemFit® AK02N 和 iMatrix-511 的无饲养层和化学定义的 ciPSC 培养系统可以稳定地维持并允许使用 N2B27 和 StemFit® AK02N 生成的 ciPSC 轻松扩增，而不会导致核型异常。ciPSCs 表达了几种多能性标记并形成了畸胎瘤，包括来自三个胚层的细胞。

更新日期：2021-06-30

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