Abstract

Cell culture techniques help us to understand the cell biology in a much better way. Transformed cell culturing will be one of the useful methods to understand the cancer cell physiology and morphology but culturing of transformed cells in-vitro is a challenging task. The cellular outcome read-outs such as cell signals, cellular secretions, gene expression and downregulation, morphology, etc., vary depending upon the microenvironment given to the cells. Therefore, it is essential to culture the cells in an environment similar to in- vivo for understanding the cell biology behind various mechanisms. This mini review aims at summarizing the conventional and advancements in culturing the transformed cells. Different techniques are highly exploited to study transformed cells, with the most commonly used 3-D semi-solid media culturing method. It is a selective media with suitable substrate that supports the growth of anchorage-independent transformed cells and is useful to elucidate results from products of each cell differentially localized in the media. Early research using soft agar helped to understand and characterize the transformed cells whereas advancements in the semi-solid media helps to screen a wide range of anti-cancer drugs, to study gene regulation, and in biosensors. The major setback of this technique remains a hindrance to extract viable cells from the media. There by to overcome the drawback of conventional semi-solid media technique, over the years, various components are incorporated and substituted to provide a within reach in-vivo conditions. More recently, using semi-solid matrix/scaffolds like matrigel/laminin/alginate, co-culturing of other cells with transformed cells provides the in vivo like niche.

Keywords: Transformed Cell Culture; Semi – Solid Media; Soft Agar; Laminin; Matrigel

References

1. Verma A. “Animal Tissue Culture”. Animal Biotechnology (2014): 211-231.
2. Borowicz S., et al. “The Soft Agar Colony Formation Assay”. Journal of Visualized Experiments 92 (2014).
3. Bielecka ZF., et al. “Hypoxic 3D in vitro culture models reveal distinct resistance processes to TKIs in renal cancer cells”. Cell and Bioscience 7 (2017): 71.
4. Hakomori SI and Murakami WT. “Glycolipids of hamster fibroblasts and derived malignant transformed cell lines”. PNAS 59.1 (1965): 254-261.
5. Pollack R., et al. “Plasminogen Activator Production Accompanies Loss of Anchorage Regulation in Transformation of Primary Rat Embryo Cells by Simian Virus 40”. Proceedings of the National Academy of Sciences 71.12 (1974): 4792-4796.
6. Bhatia S. “Introduction to animal tissue culture science”. In Introduction to Pharmaceutical Biotechnology, IOP Publishing 3 (2019).
7. Lodish H., et al. Section 6.2, Growth of Animal Cells in Culture. Molecular Cell Biology. 4th edition. New York: W. H. Freeman.
8. Kapałczyńska M., et al. “2D and 3D cell cultures - a comparison of different types of cancer cell cultures”. Archives of Medical Science 14.4 (2018): 910-919.
9. Rotem A., et al. “Alternative to the soft-agar assay that permits high-throughput drug and genetic screens for cellular transformation”. Proceedings of the National Academy of Sciences 112.18 (2015): 5708-5713.
10. Yoshida S., et al. “Stimulatory effect of reconstituted basement membrane components (matrigel) on the colony formation of a panel of human lung cancer cell lines in soft agar”. Journal of Cancer Research and Clinical Oncology 123.6 (1997): 301-309.
11. Kubota K., et al. “Comparison between Agar and Methylcellulose Cultures of Human Leukemic Cells”. Cancer Research 41 (1981): 3052-3057.
12. Freedman V. “Cellular tumorigenicity in nude mice: Correlation with cell growth in semi-solid medium”. Cell 3.4 (1974): 355-359.
13. Graham F L., et al. “Characteristics of a Human Cell Line Transformed by DNA from Human Adenovirus Type 5”. Journal of General Virology 36.1 (1977): 59-72.
14. Huschtscha L I and Holliday R. “Limited and unlimited growth of sv40- transformed cells from human diploid mrc-5 fibroblasts”. 99 (1983): 77-99.
15. Idoine JB., et al. “Ratliver cells in culture: Effect of storage, long-term culture, and transformation on some enzyme levels”. In Vitro 12.8 (1976): 541-553.
16. Horibata S., et al. “Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells”. Journal of Visualized Experiments (JoVE) (2015): e52727.
17. Khoruzhenko AI. “Optimization of tumor cell culture conditions in soft agar for subsequent immunohistochemical analysis”. Biopolymers and Cell 28.4 (2012): 302-305.
18. Bush H and Shodell M. “Cell cycle changes in transformed cells growing under serum-free conditions”. Journal of Cellular Physiology 90.3 (1977): 573-583.
19. DULBECCO R. “Topoinhibition and Serum Requirement of Transformed and Untransformed Cells”. Nature 227.5260 (1970): 802-806.
20. Kuroki T. “Colony formation of mammalian cells on agar plates and its application to Lederberg’s replica plating”. Experimental Cell Research 80.1 (1973): 55-62.
21. Freedman V H and Shin S. Brief Communication 1,2. 58.6 (2018): 1873-1875.
22. Barrett JC., et al. “Reexaminaron of the Role of Plasminogen Activator Production for Growth in Semisolid Agar of Neoplastic Hamster Cells”. 40 (1980): 1438-1443.
23. Adams A., et al. “Sensitivity of the Epstein-Barr virus transformed human lymphoid cell lines to interferon”. Journal of General Virology 28.2 (1975): 207-217.
24. Benham V., et al. “Identifying chemopreventive agents for obesity-associated cancers using an efficient, 3D high-throughput transformation assay”. Scientific Reports 9.1 (2019).
25. Vashist A., et al. “Recent advances in hydrogel based drug delivery systems for the human body”. Journal of Materials Chemistry B 2.2 (2014): 147-166.
26. Edmondson R., et al. “Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors”. ASSAY and Drug Development Technologies 12.4 (2014): 207-218.
27. Cavo M., et al. “A new cell-laden 3D Alginate-Matrigel hydrogel resembles human breast cancer cell malignant morphology, spread and invasion capability observed “in vivo”. Scientific Reports 8.1 (2018).
28. Brodaczewska K., et al. “Metastatic renal cell carcinoma cells growing in 3D on poly‑D‑lysine or laminin present a stem‑like phenotype and drug resistance”. Oncology Reports (2019).
29. Tasdemir N., et al. “Comprehensive Phenotypic Characterization of Human Invasive Lobular Carcinoma Cell Lines in 2D and 3D Cultures”. Cancer Research 78.21 (2018): 6209-6222.
30. Fukazawa H., et al. “A Microplate Assay for Quantitation of Anchorage-Independent Growth of Transformed Cells”. Analytical Biochemistry 228.1 (1995): 83-90.
31. Lovitt C., et al. “Advanced Cell Culture Techniques for Cancer Drug Discovery”. Biology 3.2 (2014): 345-367. 
32. Macpherson I. “Soft Agar Techniques”. Tissue Culture (1973): 276-280.
33. Buick N R., et al. “Application of in vitro soft agar techniques for growth of tumor cells to the study of colon cancer”. ACS Journals 45 (1980): 1238-1242.
34. Maliszewska-Olejniczak, K., et al. “Development of extracellular matrix supported 3D culture of renal cancer cells and renal cancer stem cells”. Cytotechnology 71.1 (2018): 149-163.
35. Stoker M., et al. “Anchorage and growth regulation in normal and virus-transformed cells”. International Journal of Cancer 3.5 (1968): 683-693.
36. Technical Library. Guidelines to Maintain Cultured cells | Thermo Fisher Scientific - UK. 

Citation

Citation: Pratibha M and Abirami B. “Mini Review - Semi-solid Media for Transformed Cell Culturing". Acta Scientific Medical Sciences 4.9 (2020): 145-151.

Copyright

Copyright: © 2020 Pratibha M and Abirami B. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.