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Application of Silicone Coating for Obtaining Cell Spheroids Using the Hanging Drop Method. P. 44–51

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Section: Medical and biological sciences

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UDC

57.085.2+576

DOI

10.37482/2687-1491-Z089

Authors

Svetlana Yu. Filippova* ORCID: https://orcid.org/0000-0002-4558-5896
Anastasiya O. Sitkovskaya* ORCID: https://orcid.org/0000-0002-6035-1756
Larisa N. Vashchenko* ORCID: https://orcid.org/0000-0003-2267-3460
Emma E. Kechedzhieva* ORCID: https://orcid.org/0000-0002-3051-6628
Irina R. Dashkova* ORCID: https://orcid.org/0000-0001-9688-9550
Tatʼana V. Ausheva* ORCID: https://orcid.org/0000-0002-7073-9463
Yuriy V. Przhedetskiy* ORCID: https://orcid.org/0000-0003-3976-0210
*National Medical Research Centre for Oncology (Rostov-on-Don, Russian Federation)
Corresponding author: Svetlana Filippova, address: ul. 14-ya liniya 63, Rostov-on-Don, 344037, Russian Federation; e-mail: filsv@yandex.ru

Abstract

In modern experimental oncology, 3D cell cultures have become particularly important, as they provide more relevant results compared to traditional 2D cultures. The problem of obtaining relevant cell models remains urgent for the study of breast cancer. One of the most common techniques for obtaining a 3D culture is the hanging drop method. Researchers are constantly developing its modifications to reduce variations in the shape and size of the resulting cell spheroids. One of the ways to solve this problem is to apply a hydrophobic coating to the surface of cell culture plastics. Silicones or Parafilm® laboratory films are often used for this purpose. As a result, the curvature of the drop surface increases, which leads to accelerated aggregation of cells in the centre of the drop. The purpose of this research was to evaluate the possibility of applying a coating made of the silicone elastomer SIEL 159-330 (Russia) to modify the hanging drop method. Materials and methods. We used the SIEL 159-330 coating cured at a temperature lower than that recommended by the manufacturer and investigated its cytotoxic properties as well as its effect on the formation of cell spheroids in a hanging drop. The material for the study was the BT-474 breast cancer cell line. Results. The research found that the tested elastomer has no effect on cell viability. At the same time, SIEL 159-330, compared to polystyrene, significantly reduces the time of cell aggregate formation in the lower part of the drop. In addition, cell spheroids of the breast cancer culture obtained on the SIEL 159-330 coating vary less in shape and size than spheroids obtained on the polystyrene or Parafilm coating.

Keywords

hanging drop method, BT-474 cell line, breast cancer, cell spheroid, silicone elastomer

References

1. Kit O.I., Shatova Yu.S., Novikova I.A., Vladimirova L.Yu., Ul’yanova E.P., Komova E.A., Kechedzhieva E.E. Ekspressiya P53 i BCL2 pri razlichnykh podtipakh raka molochnoy zhelezy [P53 and BCL2 Expression in Different Breast Cancer Subtypes]. Fundamental’nye issledovaniya, 2014, no. 10-1, pp. 85–88.
2. Souza A.G., Silva I.B.B., Campos-Fernandez E., Barcelos L.S., Souza J.B., Marangoni K., Goulart L.R., Alonso-Goulart V. Comparative Assay of 2D and 3D Cell Culture Models: Proliferation, Gene Expression and Anticancer Drug Response. Curr. Pharm. Des., 2018, vol. 24, no.15, pp. 1689–1694. DOI: 10.2174/1381612824666180404152304
3. Mezhevova I.V., Sitkovskaya A.O., Kit O.I. Primary Tumor Cell Cultures: Current Methods of Obtaining and Subcultivation. South Russ. J. Cancer, 2020, vol. 1, no. 3, pp. 36–49. DOI: 10.37748/2687-0533-2020-1-3-4
4. Costa E.C., Moreira A.F., de Melo-Diogo D., Gaspar V.M., Carvalho M.P., Correia I.J. 3D Tumor Spheroids: An Overview on the Tools and Techniques Used for Their Analysis. Biotechnol. Adv., 2016, vol. 34, no. 8, pp. 1427–1441. DOI: 10.1016/j.biotechadv.2016.11.002
5. Timofeeva S.V., Shamova T.V., Sitkovskaya A.O. 3D-bioprinting mikrookruzheniya opukholi: poslednie dostizheniya [3D Bioprinting of Tumor Microenvironment: Recent Achievements]. Zhurnal obshchey biologii, 2021, vol. 82, no. 5, pp. 389–400. DOI: 10.31857/s0044459621050067
6. Nunes A.S., Barros A.S., Costa E.C., Moreira A.F., Correia I.J. 3D Tumor Spheroids as in vitro Models to Mimic in vivo Human Solid Tumors Resistance to Therapeutic Drugs. Biotechnol. Bioeng., 2019, vol. 116, no. 1, pp. 206–226. DOI: 10.1002/bit.26845
7. Kelm J.M., Timmins N.E., Brown C.J., Fussenegger M., Nielsen L.K. Method for Generation of Homogeneous Multicellular Tumor Spheroids Applicable to a Wide Variety of Cell Types. Biotechnol. Bioeng., 2003, vol. 83, no. 2, pp. 173–180. DOI: 10.1002/bit.10655
8. Oliveira M.B., Neto A.I., Correia C.R., Rial-Hermida M.I., Alvarez-Lorenzo C., Mano J.F. Superhydrophobic Chips for Cell Spheroids High-Throughput Generation and Drug Screening. ACS Appl. Mater. Interfaces., 2014, vol. 6, no. 12, pp. 9488–9495. DOI: 10.1021/am5018607
9. Fu J.J., Lv X.H., Wang L.X., He X., Li Y., Yu L., Li C.M. Cutting and Bonding Parafilm® to Fast Prototyping Flexible Hanging Drop Chips for 3D Spheroid Cultures. Cell. Mol. Bioeng., 2021, vol. 14, pp. 187–199. DOI: 10.1007/s12195-020-00660-x
10. Kuo C.-T., Wang J.-Y., Lin Y.-F., Wo A.M., Chen B.P.C., Lee H. Three-Dimensional Spheroid Culture Targeting Versatile Tissue Bioassays Using a PDMS-Based Hanging Drop Array. Sci. Rep., 2017, vol. 7. Art. no. 4363. DOI: 10.1038/s41598-017-04718-1
11. Kraev I.D., Popkov O.V., Shul’deshov E.M., Sorokin A.E., Yurkov G.Yu. Perspektivy ispol’zovaniya kremniyorganicheskikh polimerov pri sozdanii sovremennykh materialov i pokrytiy razlichnykh naznacheniy [Prospects for the Use of Organosilicon Elastomers in the Development of Modern Polymer Materials and Coatings for Various Purposes]. Trudy VIAM, 2017, no. 12, pp. 48–62. DOI: 10.18577/2307-6046-2017-0-12-5-5
12. Nanush’yan S.R. Kremniyorganicheskie materialy uskorennoy vulkanizatsii: istoriya sozdaniya i razvitiya napravleniya [Fast-Curing Organosilicon Materials: History of Creation and Development]. Khimicheskaya promyshlennost’ segodnya, 2015, no. 11, pp. 21–26.
13. Guo X., Chen Y., Ji W., Chen X., Li C., Ge R. Enrichment of Cancer Stem Cells by Agarose Multi-Well Dishes and 3D Spheroid Culture. Cell Tissue Res., 2019, vol. 375, no. 2, pp. 397–408. DOI: 10.1007/s00441-018-2920-0
14. Wan L., Neumann C.A., LeDuc P.R. Tumor-on-a-Chip for Integrating a 3D Tumor Microenvironment: Chemical and Mechanical Factors. Lab. Chip, 2020, vol. 20, no. 5, pp. 873–888. DOI: 10.1039/c9lc00550a



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