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一种适用于细胞治疗方法的围产期干细胞分离新预测技术。

A New Predictive Technology for Perinatal Stem Cell Isolation Suited for Cell Therapy Approaches.

作者信息

Zia Silvia, Martini Giulia, Pizzuti Valeria, Maggio Alessia, Simonazzi Giuliana, Reschiglian Pierluigi, Bonsi Laura, Alviano Francesco, Roda Barbara, Zattoni Andrea

机构信息

Stem Sel srl., 40127 Bologna, Italy.

Unit of Histology, Department of Experimental, Diagnostic and Specialty Medicine, Embryology and Applied Biology, University of Bologna, 40126 Bologna, Italy.

出版信息

Micromachines (Basel). 2021 Jun 30;12(7):782. doi: 10.3390/mi12070782.

Abstract

The use of stem cells for regenerative applications and immunomodulatory effect is increasing. Amniotic epithelial cells (AECs) possess embryonic-like proliferation ability and multipotent differentiation potential. Despite the simple isolation procedure, inter-individual variability and different isolation steps can cause differences in isolation yield and cell proliferation ability, compromising reproducibility observations among centers and further applications. We investigated the use of a new technology as a diagnostic tool for quality control on stem cell isolation. The instrument label-free separates cells based on their physical characteristics and, thanks to a micro-camera, generates a live fractogram, the fingerprint of the sample. Eight amniotic membranes were processed by trypsin enzymatic treatment and immediately analysed. Two types of profile were generated: a monomodal and a bimodal curve. The first one represented the unsuccessful isolation with all recovered cell not attaching to the plate; while for the second type, the isolation process was successful, but we discovered that only cells in the second peak were alive and resulted adherent. We optimized a Quality Control (QC) method to define the success of AEC isolation using the fractogram generated. This predictive outcome is an interesting tool for laboratories and cell banks that isolate and cryopreserve fetal annex stem cells for research and future clinical applications.

摘要

干细胞在再生应用和免疫调节方面的应用日益增加。羊膜上皮细胞(AECs)具有类似胚胎的增殖能力和多能分化潜能。尽管分离过程简单,但个体差异和不同的分离步骤会导致分离产量和细胞增殖能力的差异,影响各中心之间观察结果的可重复性以及进一步的应用。我们研究了一种新技术作为干细胞分离质量控制的诊断工具。该仪器无需标记,根据细胞的物理特性分离细胞,并借助微型相机生成实时分形图,即样本的指纹图谱。通过胰蛋白酶酶处理对八张羊膜进行处理,并立即进行分析。生成了两种类型的图谱:单峰曲线和双峰曲线。第一种表示分离失败,所有回收的细胞都未附着在培养板上;而对于第二种类型,分离过程成功,但我们发现只有第二个峰中的细胞存活并附着。我们优化了一种质量控制(QC)方法,利用生成的分形图来确定AEC分离的成功与否。这种预测结果对于为研究和未来临床应用分离和冷冻保存胎儿附属干细胞的实验室和细胞库来说是一种有趣的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3037/8305015/f343a036ba89/micromachines-12-00782-g001.jpg

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