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在自动化封闭培养系统中制造源自人诱导多能干细胞的视网膜色素上皮细胞片用于再生医学。

Fabricating retinal pigment epithelial cell sheets derived from human induced pluripotent stem cells in an automated closed culture system for regenerative medicine.

机构信息

Center for Exploratory Research, Research & Development Group, Hitachi, Ltd., Kobe, Hyogo, Japan.

Laboratory for Retinal Regeneration, RIKEN Center for Biosystems Dynamics Research, Kobe, Hyogo, Japan.

出版信息

PLoS One. 2019 Mar 13;14(3):e0212369. doi: 10.1371/journal.pone.0212369. eCollection 2019.

Abstract

Regenerative medicine has received a lot of attention as a novel strategy for injuries and diseases that are difficult to cure using current techniques. Cell production, which is vital for regenerative medicine, has undergone remarkable progress via breakthroughs in developmental biology and tissue engineering; currently, cell production requires numerous experimental operators performing manual, small-scale cell cultures. Other major obstacles for cell production and regenerative medicine include the variable quality of products based on the experimental procedure, the skills of operators, the level of labor required for production, and costs. Technological developments are required to overcome this, including automation instead of manual culture. Age-related macular regeneration (AMD) is a refractory ocular disease that causes severe deterioration in central vision due to senescence in the retinal pigment epithelium (RPE). Recently, we performed an autologous transplantation of induced pluripotent stem (iPS) cell-derived RPE cell sheets and started clinical research on allografts from RPE cell suspensions differentiated from iPS cells. The use of regenerative therapies for AMD using iPS cell-derived RPE is expected to become more widespread. In the present study, human iPS cell-derived RPE cells were cultured to form RPE cell sheets using equipment with a closed culture module. The quality of the automated cultured RPE cell sheets was confirmed by comparing their morphological and biological properties with those of manually generated RPE cell sheets. As a result, machine-cultured RPE sheets displayed the same quality as manually cultured RPE sheets, showing that iPS cell-derived RPE cell sheets were successfully cultured by an automated process.

摘要

再生医学作为一种治疗目前技术难以治愈的损伤和疾病的新策略,受到了广泛关注。通过在发育生物学和组织工程方面的突破,再生医学所必需的细胞生产取得了显著进展;目前,细胞生产需要许多实验操作人员进行手动、小规模的细胞培养。细胞生产和再生医学的其他主要障碍包括基于实验程序、操作人员技能、生产所需劳动力水平和成本的产品质量的可变性。需要进行技术开发来克服这些障碍,包括用自动化代替手动培养。年龄相关性黄斑变性(AMD)是一种难治性眼部疾病,由于视网膜色素上皮(RPE)衰老,导致中央视力严重恶化。最近,我们进行了自体诱导多能干细胞(iPS)细胞衍生的 RPE 细胞片的移植,并开始了异体 RPE 细胞悬浮液从 iPS 细胞分化而来的临床研究。使用 iPS 细胞衍生的 RPE 进行 AMD 的再生疗法有望得到更广泛的应用。在本研究中,使用具有封闭培养模块的设备培养人 iPS 细胞衍生的 RPE 细胞,以形成 RPE 细胞片。通过比较其形态和生物学特性与手动生成的 RPE 细胞片,确认了自动化培养的 RPE 细胞片的质量。结果表明,机器培养的 RPE 片与手动培养的 RPE 片具有相同的质量,表明 iPS 细胞衍生的 RPE 细胞片已成功通过自动化过程培养。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0c1a/6415881/ba9ad60374f7/pone.0212369.g001.jpg

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