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去端胶原蛋白支持人诱导多能干细胞的三维培养。

Atelocollagen supports three-dimensional culture of human induced pluripotent stem cells.

作者信息

Nakashima Yoshiki, Tsukahara Masayoshi

机构信息

CiRA Foundation, Research and Development Center, Nakanoshima Qross, Osaka 530-005, Japan.

出版信息

Mol Ther Methods Clin Dev. 2024 Jul 20;32(3):101302. doi: 10.1016/j.omtm.2024.101302. eCollection 2024 Sep 12.

Abstract

As autologous induced pluripotent stem cell (iPSC) therapy requires a custom-made small-lot cell production line, and the cell production method differs significantly from the existing processes for producing allogeneic iPSC stocks for clinical use. Specifically, mass culture to produce stock is no longer necessary; instead, a series of operations from iPSC production to induction of differentiation of therapeutic cells must be performed continuously. A three-dimensional (3D) culture method using small, closed-cell manufacturing devices is suitable for autologous iPSC therapy. The use of such devices avoids the need to handle many patient-derived specimens in a single clean room; handling of cell cultures in an open system in a cell processing facility increases the risk of infection. In this study, atelocollagen beads were evaluated as a 3D biomaterial to assist 3D culture in the establishment, expansion culture, and induction of differentiation of iPSCs. It was found that iPSCs can be handled in a closed-cell device with the same ease as use of a two-dimensional (2D) culture when laminin-511 is added to the medium. In conclusion, atelocollagen beads enable 3D culture of iPSCs, and the quality of the obtained cells is at the same level as those derived from 2D culture.

摘要

由于自体诱导多能干细胞(iPSC)疗法需要定制的小批量细胞生产线,且细胞生产方法与现有的临床用异基因iPSC库生产流程有显著差异。具体而言,不再需要进行大规模培养来生产库存;相反,必须连续进行从iPSC生产到治疗性细胞分化诱导的一系列操作。使用小型、封闭细胞制造装置的三维(3D)培养方法适用于自体iPSC疗法。使用此类装置可避免在单个洁净室内处理许多患者来源的样本;在细胞处理设施的开放系统中处理细胞培养物会增加感染风险。在本研究中,对去端肽胶原蛋白珠作为3D生物材料进行了评估,以协助iPSC的建立、扩增培养和分化诱导过程中的3D培养。结果发现,当在培养基中添加层粘连蛋白-511时,iPSC可以在封闭细胞装置中处理,其操作简便程度与二维(2D)培养相同。总之,去端肽胶原蛋白珠能够实现iPSC的3D培养,且所获得细胞的质量与2D培养来源的细胞质量处于同一水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cfd0/11342089/7b6c473f5426/fx1.jpg

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