iPS細胞から広がる医療の未来

霊長類ES/iPS細胞用培地 (Primate ES Cell Medium) プロトコル

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1. ヒトiPS細胞の形態

iPS細胞の形態について解説します。細胞の状態の判断にご活用ください。良い状態のiPS細胞は、コロニーの輪郭が明瞭で内部の細胞密度が高くなっています。

iPS細胞の培養に使用するフィーダー細胞は、その播種密度が重要になります。3×10⁴細胞/cm²が目安になります。

Dissociation Solution for human ES/iPS Cells (CTK溶液) を用いることで、高生存率かつ簡便な継代が可能になります。ピペッティング操作だけで、適度な大きさのコロニーにほぐれ、そのまま継代できます。通常3－4日に1回、継代を行います。

Freezing Medium for human ES/iPS Cellsを使用し、ガラス化法で凍結することで、高い生存率での凍結保存が可能になります。ガラス化法とは、細胞を液体窒素で急速凍結する方法で、細胞へのダメージを最小限に抑えます。また、融解時も、温めておいた培養液を凍結細胞に直接加えることで急速解凍を行います。凍結・融解の作業に時間がかかると、生存率が低下するのでご注意ください。

霊長類ES/iPS細胞用培地 (Primate ES Cell Medium) を実験にご使用いただいた論文のリストはこちらです。

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• Nasu, Akira, et al. “Genetically matched human iPS cells reveal that propensity for cartilage and bone differentiation differs with clones, not cell type of origin.” PloS one 8.1 (2013): e53771.
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• Tanaka, Akihito, et al. “Efficient and reproducible myogenic differentiation from human iPS cells: prospects for modeling Miyoshi Myopathy in vitro.” PloS one 8.4 (2013): e61540.
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• Matsuura, Katsuhisa, et al. “Creation of human cardiac cell sheets using pluripotent stem cells.” Biochemical and biophysical research communications 425.2 (2012): 321-327.
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• Iwabuchi, Kumiko A., et al. “ECAT11/L1td1 Is Enriched in ESCs and Rapidly Activated During iPSCGeneration, but It Is Dispensable for the Maintenance and Induction of Pluripotency.” PloS one 6.5 (2011): e20461.
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• Ogawa, Shin-ichiro, et al. “Induction of oligodendrocyte differentiation from adult human fibroblast-derived induced pluripotent stem cells.” In Vitro Cellular & Developmental Biology-Animal 47.7 (2011): 464-469.
• Okita, Keisuke, et al. “A more efficient method to generate integration-free human iPS cells.” Nature methods 8.5 (2011): 409-412.
• Takata, Akemi, et al. “Direct differentiation of hepatic cells from human induced pluripotent stem cells using a limited number of cytokines.” Hepatology international 5.4 (2011): 890-898.
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• Yahata, Naoki, et al. “Anti-Aβ drug screening platform using human iPS cell-derived neurons for the treatment of Alzheimer’s disease.” PLoS One 6.9 (2011): e25788.
• Fujioka, Tsuyoshi, et al. “Establishment of induced pluripotent stem cells from human neonatal tissues.” Human cell 23.3 (2010): 113-118.
• Kazuki, Yasuhiro, et al. “Complete genetic correction of ips cells from Duchenne muscular dystrophy.” Molecular Therapy 18.2 (2010): 386-393.
• Lee, Tae-Hee, et al. “Functional recapitulation of smooth muscle cells via induced pluripotent stem cells from human aortic smooth muscle cells.” Circulation research 106.1 (2010): 120-128.
• Miyoshi, Keiko, et al. “Generation of human induced pluripotent stem cells from oral mucosa.” Journal of bioscience and bioengineering 110.3 (2010): 345-350.
• Otsuji, Tomomi G., et al. “Progressive maturation in contracting cardiomyocytes derived from human embryonic stem cells: Qualitative effects on electrophysiological responses to drugs.” Stem cell research 4.3 (2010): 201-213.
• Sakurai, Kenji, et al. “Efficient integration of transgenes into a defined locus in human embryonic stem cells.” Nucleic acids research 38.7 (2010): e96-e96.
• Tamaoki, N, et al. “Dental pulp cells for induced pluripotent stem cell banking.” Journal of dental research 89.8 (2010): 773-778.
• Oka, Y., et al. “293FT cells transduced with four transcription factors (OCT4, SOX2, NANOG, and LIN28) generate aberrant ES-like cells.” J Stem cell Regenerative Med 3 (2010): 149-56.
• Tamaoki, N., et al. “Dental pulp cells for induced pluripotent stem cell banking.”Journal of dental research 89.8 (2010): 773-778.
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