Nishiyama Yuichiro, Iwanami Akio, Kohyama Jun, Itakura Go, Kawabata Soya, Sugai Keiko, Nishimura Soraya, Kashiwagi Rei, Yasutake Kaori, Isoda Miho, Matsumoto Morio, Nakamura Masaya, Okano Hideyuki
Department of Orthopedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan; Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
Department of Orthopedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
Neurosci Res. 2016 Jun;107:20-9. doi: 10.1016/j.neures.2015.11.011. Epub 2016 Jan 22.
Stem cells represent a potential cellular resource in the development of regenerative medicine approaches to the treatment of pathologies in which specific cells are degenerated or damaged by genetic abnormality, disease, or injury. Securing sufficient supplies of cells suited to the demands of cell transplantation, however, remains challenging, and the establishment of safe and efficient cell banking procedures is an important goal. Cryopreservation allows the storage of stem cells for prolonged time periods while maintaining them in adequate condition for use in clinical settings. Conventional cryopreservation systems include slow-freezing and vitrification both have advantages and disadvantages in terms of cell viability and/or scalability. In the present study, we developed an advanced slow-freezing technique using a programmed freezer with a magnetic field called Cells Alive System (CAS) and examined its effectiveness on human induced pluripotent stem cell-derived neural stem/progenitor cells (hiPSC-NS/PCs). This system significantly increased cell viability after thawing and had less impact on cellular proliferation and differentiation. We further found that frozen-thawed hiPSC-NS/PCs were comparable with non-frozen ones at the transcriptome level. Given these findings, we suggest that the CAS is useful for hiPSC-NS/PCs banking for clinical uses involving neural disorders and may open new avenues for future regenerative medicine.
干细胞是再生医学治疗特定细胞因基因异常、疾病或损伤而退化或受损的病理状况时潜在的细胞资源。然而,确保获得足够数量适合细胞移植需求的细胞仍然具有挑战性,建立安全有效的细胞库程序是一个重要目标。冷冻保存可使干细胞长时间储存,同时保持其在临床应用中的适宜状态。传统的冷冻保存系统包括慢速冷冻和玻璃化,在细胞活力和/或可扩展性方面都有优缺点。在本研究中,我们开发了一种先进的慢速冷冻技术,使用带有磁场的程序冷冻器,即细胞存活系统(CAS),并研究了其对人诱导多能干细胞来源的神经干细胞/祖细胞(hiPSC-NS/PCs)的有效性。该系统显著提高了复苏后的细胞活力,对细胞增殖和分化的影响较小。我们进一步发现,冻融后的hiPSC-NS/PCs在转录组水平上与未冷冻的细胞相当。基于这些发现,我们认为CAS对于涉及神经疾病的临床应用中hiPSC-NS/PCs的细胞库构建很有用,并可能为未来的再生医学开辟新途径。