1 Department of Biomedical Engineering, Cleveland Clinic, Cleveland, OH, USA.
2 Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH, USA.
SLAS Technol. 2017 Apr;22(2):217-223. doi: 10.1177/2211068216676587. Epub 2016 Nov 11.
Stem and progenitor cells derived from human tissues are being developed as cell sources for cell-based assays and therapies. However, tissue-derived stem and progenitor cells are heterogeneous. Differences in observed clones of stem cells likely reflect important aspects of the underlying state of the source cells, as well as future potency for cell therapies. This paper describes a colony analysis and picking device that provides quantitative analysis of heterogeneous cell populations and precise tools for cell picking for research or biomanufacturing applications. We describe an integrated robotic system that enables image acquisition and automated image analysis to be coupled with rapid automated selection of individual colonies in adherent cell cultures. Other automated systems have demonstrated feasibility with picking from semisolid media or off feeder layers. We demonstrate the capability to pick adherent bone-derived stem cells from tissue culture plastic. Cells are efficiently picked from a target site and transferred to a recipient well plate. Cells demonstrate viability and adherence and maintain biologic potential for surface markers CD73 and CD90 based on phase contrast and fluorescence imaging 6 days after transfer. Methods developed here can be applied to the study of other stem cell types and automated culture of cells.
从人体组织中提取的干细胞和祖细胞被开发为基于细胞的检测和治疗的细胞来源。然而，组织来源的干细胞和祖细胞是异质的。观察到的干细胞克隆的差异可能反映了来源细胞的潜在状态的重要方面，以及细胞治疗的未来潜力。本文描述了一种集落分析和挑取装置，该装置可对异质细胞群进行定量分析，并为研究或生物制造应用提供精确的细胞挑取工具。我们描述了一种集成的机器人系统，该系统能够将图像采集和自动图像分析与在贴壁细胞培养物中快速自动选择单个集落相结合。其他自动化系统已经证明了从半固体培养基或无饲养层中挑取的可行性。我们展示了从组织培养塑料中挑取贴壁骨源性干细胞的能力。细胞从目标部位被高效地挑取，并转移到接受孔板中。基于相差和荧光成像，转移后 6 天，细胞显示出活力和黏附性，并保持表面标志物 CD73 和 CD90 的生物学潜力。这里开发的方法可应用于其他干细胞类型的研究和细胞的自动化培养。
