ISSN 1866-8836
Клеточная терапия и трансплантация

BS-07. Development of magnetic nanoscale systems suitable for clinical selection of therapeutically relevant populations of human cells

Timofey A. Pyatiizbyantsev1, Aya Darwish2, Olga S. Epifanovskaya1, Alexander S. Timin2, Ksenia A. Mitusova2, Alena I. Shakirova1, Kirill V. Lepik1

1 RM Gorbacheva Research Institute, Pavlov University, St. Petersburg, Russia
2 Peter the Great St. Petersburg Polytechnic University, St. Petersburg, Russia


Contact: Timofey A. Pyatiizbyantsev, e-mail: tim.fivehuts@gmail.com

doi 10.18620/ctt-1866-8836-2023-12-3-1-176

Summary

In modern research and clinical practice, there is a need for development of new technologies and materials that can provide more accurate and efficient selection of therapeutically relevant populations of human cells, such as T-lymphocytes and hematopoietic stem cells (HSCs). Magnetic selection is among the most effective methods of cell separation [Polyakova, et al., 2023]. This method is based on the use of ferromagnetic particles coated with antibodies specific to the therapeutic cell populations. This approach allows for the isolation of cells directly from biological fluids (blood, bone marrow, etc.) without the additional labor associated with removing magnetic carriers from the cell suspension. Our objective was to assess toxicity and feasibility for magnetic selection of human cells of the developed magnetic nanoscale systems, including magnetoelectric nanoparticle (MENs) and magnetoelectric multilayer capsules (CAPs).

Materials and methods

Toxicity of the developed MENs [Stimphil N, et al. 2017] and CAPs [Koshev N, et al. 2023] was investigated on model cell lines K-562 and THP-1, as well as on primary peripheral blood mononuclear cells (PBMCs). Cells at a concentration of 1×106 cells/mL were incubated with MENs and CAPs at a wide range of concentrations (from 0.1 to 50 µM) for 12 hours. Cell viability was assessed by flow cytometry using Annexin V-FITC/7-AAD staining (BioLegend, USA), as well as by AlamarBlue analysis (Resazurin test, Sigma-Aldrich, USA). The efficiency of magnetic selection using MENs and CAPs without prior conjugation with antibodies was evaluated in a model cell line of human monocytes (THP-1), which shows high phagocytic capacity. The selection was performed on LS Columns (Miltenyi Biotec, Germany) after incubating cells with MENs and CAPs overnight.

Results

Cell viability after incubation with MENs and CAPs at substance concentrations ranging from 0.1 µM to 50 µM per 1×106 cells showed an exponential decrease, thus allowing definition of optimal concentrations of MENs and CAPs for maintaining the close-to-control viability of cells, i.e., 1 µM for both variants of the particles. Application of magnetic selection technique has demonstrated that, after a single incubation of the THP-1 cell line with MENs and CAPs at the optimal concentration followed by standard washing at the magnetic column, the non-specific interaction of nanoscale magnetic particles and capsules with phagocytic THP-1 cells resulted in effective selection of 41.0% and 41.7% of cells for MENs and CAPs, respectively.

Conclusions

We have determined optimal concentrations of magnetic nanoscale systems, i.e., particles and capsules, for magnetic selection. The results of toxicity tests showed high cell line survival rates at the maximal concentration, thus suggesting a potential for usage of such systems for magnetic selection. However, further research is needed to increase the efficiency of magnetic selection and to improve specificity of the particle interaction with therapeutically relevant populations of human cells.

Acknowledgement

MEN and CAP synthesis experiments were conducted under the financial support of the Ministry of science and higher education of Russian Federation (FSEG-2022-0012).

Keywords

Magnetic nanoscale systems, selection, T-lymphocytes, HSC, peripheral blood mononuclear cells.


Supplement 12-3
09/30/2023

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doi 10.18620/ctt-1866-8836-2023-12-3-1-176

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