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.

Highlights

  • Advanced therapy medicinal products (ATMP) are medicines based on genes, cells and tissues to treat human diseases

  • No difference was observed between the use of different trypsin concentration; both concentrations showed the same predictivity about the isolation protocol

  • This study evidenced the potential of this technology as a quality control system to predict the outcome of stem cell isolation procedure and the quality of freshly isolated predict the outcome of stem cell isolation procedure and the quality of freshly isolated cells, cells, amniotic epithelial (AECs)

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Summary

Introduction

Advanced therapy medicinal products (ATMP) are medicines based on genes, cells and tissues to treat human diseases. The somatic-cell therapy consists of cells infusion that replaces tissue functions, cures and prevents diseases. Adult SCs are widely used to treat malignant diseases like leukemia by hematopoietic stem cells transplantation, and Graft Versus Host Disease (GVHD) by bone marrow mesenchymal stem cells (BMMSCs) for their immunomodulatory capacity [1,2,3,4,5]. Perinatal SCs have gained attention because they possess wide differentiation potential and tolerogenic ability [6]. Placenta is a rich source of stem cells: mesenchymal and epithelial cells with staminal characteristics can be derived and it was proven their therapeutic potential in various disease models [7].

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