İki-boyutlu ve Üç-boyutlu Hücre Kültür Modellerinde Güncel Gelişmeler ve Uygulamalar

Güneş Kibar; Atakan Tevlek

doi:10.17827/aktd.1607163

Derleme

Current Developments and Applications in 2D and 3D Cell Culture Models

Yıl 2025, Cilt: 34 Sayı: 2, 119 - 128, 30.06.2025

Güneş Kibar , Atakan Tevlek

https://doi.org/10.17827/aktd.1607163

Öz

Cell culture models are critical tools in basic and applied biomedical research. While traditional 2B cell culture systems have been widely used due to their simplicity and ease of application, they fall short in mimicking the natural microenvironment of cells. To overcome these limitations, 3D cell culture models have been developed to better replicate cell-cell and cell-matrix interactions, enabling more accurate modeling of biological processes. In particular, microfluidic-based systems, organoids, and biomaterial-enriched 3D platforms have paved the way for innovative applications ranging from cancer research to tissue engineering. This review explores the historical development, advantages, disadvantages, and current applications of 2D and 3D cell culture models, along with their potential in biomedical research. Additionally, the impact of advanced technologies such as dynamic and magnetic cell culture on 3D models is discussed, offering a perspective on innovative approaches in this field.

Anahtar Kelimeler

Tissue engineering, organoids, microfluidic systems, magnetic cell culture

Proje Numarası

123M943

Kaynakça

1. Ramezankhani, R., et al., Organoid and microfluidics-based platforms for drug screening in COVID-19. Drug Discovery Today, 2022. 27(4): p. 1062-1076.
2. Deb, B., H. Shah, and S. Goel, Current global vaccine and drug efforts against COVID-19: Pros and cons of bypassing animal trials. Journal of biosciences, 2020. 45: p. 1-10.
3. Maeda, H. and M. Khatami, Analyses of repeated failures in cancer therapy for solid tumors: poor tumor-selective drug delivery, low therapeutic efficacy and unsustainable costs. Clinical and translational medicine, 2018. 7: p. 1-20.
4. Shaverdian, N., et al., Effects of tumor mutational burden and gene alterations associated with radiation response on outcomes of postoperative radiation therapy in non-small cell lung cancer. International Journal of Radiation Oncology* Biology* Physics, 2022. 113(2): p. 335-344.
5. Mirfakhraie, R., et al., Treatment failure in acute myeloid leukemia: focus on the role of extracellular vesicles. Leukemia Research, 2022. 112: p. 106751.
6. Di Federico, A., et al., Immunotherapy in pancreatic cancer: why do we keep failing? A focus on tumor immune microenvironment, predictive biomarkers and treatment outcomes. Cancers, 2022. 14(10): p. 2429.
7. Hudu, S.A., et al., Cell culture, technology: enhancing the culture of diagnosing human diseases. Journal of clinical and diagnostic research: JCDR, 2016. 10(3): p. DE01.
8. Huang, D., et al., Liver spheroids on chips as emerging platforms for drug screening. Engineered Regeneration, 2022.
9. Barbosa, M.A., et al., 3B cell culture models as recapitulators of the tumor microenvironment for the screening of anti-cancer drugs. Cancers, 2022. 14(1): p. 190.
10. Marchini, A. and F. Gelain, Synthetic scaffolds for 3B cell cultures and organoids: applications in regenerative medicine. Critical reviews in biotechnology, 2022. 42(3): p. 468-486.
11. Fontoura, J.C., et al., Comparison of 2B and 3B cell culture models for cell growth, gene expression and drug resistance. Materials Science and Engineering: C, 2020. 107: p. 110264.
12. Namekawa, T., et al., Application of prostate cancer models for preclinical study: advantages and limitations of cell lines, patient-derived xenografts, and three-dimensional culture of patient-derived cells. Cells, 2019. 8(1): p. 74.
13. Kapałczyńska, M., et al., 2B and 3B cell cultures–a comparison of different types of cancer cell cultures. Archives of Medical Science, 2018. 14(4): p. 910-919.
14. Mittal, R., et al., Organ‐on‐chip models: implications in drug discovery and clinical applications. Journal of cellular physiology, 2019. 234(6): p. 8352-8380.
15. Duval, K., et al., Modeling physiological events in 2B vs. 3B cell culture. Physiology, 2017. 32(4): p. 266-277.
16. Harrison, R. Biology and medicine. Observations on the living developing nerve fiber. in Sci. Proc. 1907.
17. LaBarbera, D.V., B.G. Reid, and B.H. Yoo, The multicellular tumor spheroid model for high-throughput cancer drug discovery. Expert opinion on drug discovery, 2012. 7(9): p. 819-830.
18. Brancato, V., et al., Could 3B models of cancer enhance drug screening? Biomaterials, 2020. 232: p. 119744.
19. Godugu, C., et al., AlgiMatrix™ based 3B cell culture system as an in-vitro tumor model for anticancer studies. PloS one, 2013. 8(1): p. e53708.
20. Marques, I.A., et al., Magnetic-Based Human Tissue 3B Cell Culture: A Systematic Review. International Journal of Molecular Sciences, 2022. 23(20): p. 12681.
21. Moro, L.G., et al., A Brief Review on the Cell Culture History: From Harrison to Organs-on-a-Chip. 2024.
22. Kibar, G., et al., Evaluation of drug carrier hepatotoxicity using primary cell culture models. Nanomedicine: Nanotechnology, Biology and Medicine, 2023. 48: p. 102651.
23. Bissell, M.J. and W.C. Hines, Why don't we get more cancer? A proposed role of the microenvironment in restraining cancer progression. Nature medicine, 2011. 17(3): p. 320-329.
24. Jensen, C. and Y. Teng, Is it time to start transitioning from 2B to 3B cell culture? Frontiers in molecular biosciences, 2020. 7: p. 33.
25. Ravi, M., et al., 3B cell culture systems: advantages and applications. Journal of cellular physiology, 2015. 230(1): p. 16-26.
26. Luckert, C., et al., Comparative analysis of 3B culture methods on human HepG2 cells. Archives of toxicology, 2017. 91: p. 393-406.
27. Aisenbrey, E.A. and W.L. Murphy, Synthetic alternatives to Matrigel. Nature Reviews Materials, 2020. 5(7): p. 539-551.
28. Carletti, E., A. Motta, and C. Migliaresi, Scaffolds for tissue engineering and 3B cell culture. 3B Cell Culture: Methods and Protocols, 2011: p. 17-39.
29. Hoarau-Véchot, J., et al., Halfway between 2B and animal models: are 3B cultures the ideal tool to study cancer-microenvironment interactions? International journal of molecular sciences, 2018. 19(1): p. 181.
30. Alghuwainem, A., A.T. Alshareeda, and B. Alsowayan, Scaffold-free 3-D cell sheet technique bridges the gap between 2-D cell culture and animal models. International journal of molecular sciences, 2019. 20(19): p. 4926.
31. Timmins, N.E. and L.K. Nielsen, Generation of multicellular tumor spheroids by the hanging-drop method. Tissue engineering, 2007: p. 141-151.
32. Amaral, R.L., et al., Comparative analysis of 3B bladder tumor spheroids obtained by forced floating and hanging drop methods for drug screening. Frontiers in physiology, 2017. 8: p. 605.
33. Barrila, J., et al., Organotypic 3B cell culture models: using the rotating wall vessel to study host–pathogen interactions. Nature Reviews Microbiology, 2010. 8(11): p. 791-801.
34. Basu, A., et al., Ready to go 3B? A semi-automated protocol for microwell spheroid arrays to increase scalability and throughput of 3B cell culture testing. Toxicology Mechanisms and Methods, 2020. 30(8): p. 590- 604.
35. Jorgensen, C. and M. Simon, In vitro human joint models combining advanced 3B cell culture and cutting- edge 3B bioprinting technologies. Cells, 2021. 10(3): p. 596.
36. Russo, M., C.M. Cejas, and G. Pitingolo, Advances in microfluidic 3B cell culture for preclinical drug development. Progress in Molecular Biology and Translational Science, 2022. 187(1): p. 163-204.
37. Li, L., et al., A microfluidic 3B hepatocyte chip for hepatotoxicity testing of nanoparticles. Nanomedicine, 2019. 14(16): p. 2209-2226.
38. Van Duinen, V., et al., Microfluidic 3B cell culture: from tools to tissue models. Current opinion in biotechnology, 2015. 35: p. 118-126.
39. Caleffi, J.T., et al., Magnetic 3B cell culture: State of the art and current advances. Life Sciences, 2021. 286: p. 120028.
40. Anil‐Inevi, M., et al., Magnetic levitation assisted biofabrication, culture, and manipulation of 3B cellular structures using a ring magnet based setup. Biotechnology and Bioengineering, 2021. 118(12): p. 4771-4785.
41. Bonnier, F., et al., Cell viability assessment using the Alamar blue assay: a comparison of 2B and 3B cell culture models. Toxicology in vitro, 2015. 29(1): p. 124-131.
42. Soares, C.P., et al., 2B and 3B-organized cardiac cells shows differences in cellular morphology, adhesion junctions, presence of myofibrils and protein expression. PloS one, 2012. 7(5): p. e38147.
43. Rodríguez-Hernández, M.A., et al., Differential effectiveness of tyrosine kinase inhibitors in 2B/3B culture according to cell differentiation, p53 status and mitochondrial respiration in liver cancer cells. Cell Death & Disease, 2020. 11(5): p. 339.
44. Luca, A.C., et al., Impact of the 3B microenvironment on phenotype, gene expression, and EGFR inhibition of colorectal cancer cell lines. PloS one, 2013. 8(3): p. e59689.
45. Liu, H., J. Lin, and K. Roy, Effect of 3B scaffold and dynamic culture condition on the global gene expression profile of mouse embryonic stem cells. Biomaterials, 2006. 27(36): p. 5978-5989.
46. Baharvand, H., et al., Differentiation of human embryonic stem cells into hepatocytes in 2B and 3B culture systems in vitro. International journal of developmental biology, 2004. 50(7): p. 645-652.
47. Padmalayam, I. and M.J. Suto, 3B cell cultures: Mimicking in vivo tissues for improved predictability in drug discovery. Annual reports in medicinal chemistry, 2012. 47: p. 367-378.
48. Breslin, S. and L. O'Driscoll, The relevance of using 3B cell cultures, in addition to 2B monolayer cultures, when evaluating breast cancer drug sensitivity and resistance. Oncotarget, 2016. 7(29): p. 45745-45756.
49. Souza, A.G., et al., Comparative assay of 2B and 3B cell culture models: proliferation, gene expression and anticancer drug response. Current pharmaceutical design, 2018. 24(15): p. 1689-1694.
50. Imamura, Y., et al., Comparison of 2B-and 3B-culture models as drug-testing platforms in breast cancer. Oncology reports, 2015. 33(4): p. 1837-1843.
51. Melissaridou, S., et al., The effect of 2B and 3B cell cultures on treatment response, EMT profile and stem cell features in head and neck cancer. Cancer cell international, 2019. 19(1): p. 1-10.
52. Wendt, D., et al., Potential and bottlenecks of bioreactors in 3B cell culture and tissue manufacturing. Advanced materials, 2009. 21(32‐33): p. 3352-3367.

İki-boyutlu ve Üç-boyutlu Hücre Kültür Modellerinde Güncel Gelişmeler ve Uygulamalar

Yıl 2025, Cilt: 34 Sayı: 2, 119 - 128, 30.06.2025

Güneş Kibar , Atakan Tevlek

https://doi.org/10.17827/aktd.1607163

Öz

Hücre kültür modelleri, temel ve uygulamalı biyomedikal araştırmalarda kritik bir araç olarak kullanılmaktadır. Geleneksel 2-boyutlu (2B) hücre kültür sistemleri, basitlikleri ve kolay uygulanabilirlikleri nedeniyle uzun yıllardır tercih edilmekle birlikte, hücrelerin doğal mikroçevrelerini yeterince taklit edememesi nedeniyle bazı sınırlamalara sahiptir. Bu eksikliklerin giderilmesi amacıyla geliştirilen 3-boyutlu (3B) hücre kültür modelleri, hücre-hücre ve hücre-matriks etkileşimlerini daha iyi yansıtarak biyolojik süreçlerin daha doğru bir şekilde modellenmesine olanak sağlamaktadır. Özellikle mikroakışkan tabanlı sistemler, organoidler ve biyomalzemelerle zenginleştirilmiş 3B platformlar, kanser araştırmalarından doku mühendisliğine kadar geniş bir yelpazede yenilikçi uygulamalara kapı aralamıştır. Bu derleme, 2B ve 3B hücre kültür modellerinin tarihsel gelişimini, avantaj ve dezavantajlarını, güncel uygulamalarını ve biyomedikal araştırmalardaki gelecekteki potansiyellerini ele almaktadır. Ayrıca, dinamik ve manyetik hücre kültürü gibi ileri teknolojilerin 3B modeller üzerindeki etkileri tartışılarak, bu alandaki yenilikçi yaklaşımlar için bir perspektif sunulmaktadır.

Anahtar Kelimeler

doku mühendisliği, organoidler, mikroakışkan sistemler, manyetik hücre kültürü

Destekleyen Kurum

TUBITAK

Proje Numarası

123M943

Teşekkür

Bu çalışma, Türkiye Bilimsel ve Teknolojik Araştırma Kurumu (TÜBİTAK) tarafından 123M943 numaralı proje ile desteklenmiştir. Projeye verdiği destekten ötürü TÜBİTAK’a teşekkürlerimizi sunarız.

Kaynakça

1. Ramezankhani, R., et al., Organoid and microfluidics-based platforms for drug screening in COVID-19. Drug Discovery Today, 2022. 27(4): p. 1062-1076.
2. Deb, B., H. Shah, and S. Goel, Current global vaccine and drug efforts against COVID-19: Pros and cons of bypassing animal trials. Journal of biosciences, 2020. 45: p. 1-10.
3. Maeda, H. and M. Khatami, Analyses of repeated failures in cancer therapy for solid tumors: poor tumor-selective drug delivery, low therapeutic efficacy and unsustainable costs. Clinical and translational medicine, 2018. 7: p. 1-20.
4. Shaverdian, N., et al., Effects of tumor mutational burden and gene alterations associated with radiation response on outcomes of postoperative radiation therapy in non-small cell lung cancer. International Journal of Radiation Oncology* Biology* Physics, 2022. 113(2): p. 335-344.
5. Mirfakhraie, R., et al., Treatment failure in acute myeloid leukemia: focus on the role of extracellular vesicles. Leukemia Research, 2022. 112: p. 106751.
6. Di Federico, A., et al., Immunotherapy in pancreatic cancer: why do we keep failing? A focus on tumor immune microenvironment, predictive biomarkers and treatment outcomes. Cancers, 2022. 14(10): p. 2429.
7. Hudu, S.A., et al., Cell culture, technology: enhancing the culture of diagnosing human diseases. Journal of clinical and diagnostic research: JCDR, 2016. 10(3): p. DE01.
8. Huang, D., et al., Liver spheroids on chips as emerging platforms for drug screening. Engineered Regeneration, 2022.
9. Barbosa, M.A., et al., 3B cell culture models as recapitulators of the tumor microenvironment for the screening of anti-cancer drugs. Cancers, 2022. 14(1): p. 190.
10. Marchini, A. and F. Gelain, Synthetic scaffolds for 3B cell cultures and organoids: applications in regenerative medicine. Critical reviews in biotechnology, 2022. 42(3): p. 468-486.
11. Fontoura, J.C., et al., Comparison of 2B and 3B cell culture models for cell growth, gene expression and drug resistance. Materials Science and Engineering: C, 2020. 107: p. 110264.
12. Namekawa, T., et al., Application of prostate cancer models for preclinical study: advantages and limitations of cell lines, patient-derived xenografts, and three-dimensional culture of patient-derived cells. Cells, 2019. 8(1): p. 74.
13. Kapałczyńska, M., et al., 2B and 3B cell cultures–a comparison of different types of cancer cell cultures. Archives of Medical Science, 2018. 14(4): p. 910-919.
14. Mittal, R., et al., Organ‐on‐chip models: implications in drug discovery and clinical applications. Journal of cellular physiology, 2019. 234(6): p. 8352-8380.
15. Duval, K., et al., Modeling physiological events in 2B vs. 3B cell culture. Physiology, 2017. 32(4): p. 266-277.
16. Harrison, R. Biology and medicine. Observations on the living developing nerve fiber. in Sci. Proc. 1907.
17. LaBarbera, D.V., B.G. Reid, and B.H. Yoo, The multicellular tumor spheroid model for high-throughput cancer drug discovery. Expert opinion on drug discovery, 2012. 7(9): p. 819-830.
18. Brancato, V., et al., Could 3B models of cancer enhance drug screening? Biomaterials, 2020. 232: p. 119744.
19. Godugu, C., et al., AlgiMatrix™ based 3B cell culture system as an in-vitro tumor model for anticancer studies. PloS one, 2013. 8(1): p. e53708.
20. Marques, I.A., et al., Magnetic-Based Human Tissue 3B Cell Culture: A Systematic Review. International Journal of Molecular Sciences, 2022. 23(20): p. 12681.
21. Moro, L.G., et al., A Brief Review on the Cell Culture History: From Harrison to Organs-on-a-Chip. 2024.
22. Kibar, G., et al., Evaluation of drug carrier hepatotoxicity using primary cell culture models. Nanomedicine: Nanotechnology, Biology and Medicine, 2023. 48: p. 102651.
23. Bissell, M.J. and W.C. Hines, Why don't we get more cancer? A proposed role of the microenvironment in restraining cancer progression. Nature medicine, 2011. 17(3): p. 320-329.
24. Jensen, C. and Y. Teng, Is it time to start transitioning from 2B to 3B cell culture? Frontiers in molecular biosciences, 2020. 7: p. 33.
25. Ravi, M., et al., 3B cell culture systems: advantages and applications. Journal of cellular physiology, 2015. 230(1): p. 16-26.
26. Luckert, C., et al., Comparative analysis of 3B culture methods on human HepG2 cells. Archives of toxicology, 2017. 91: p. 393-406.
27. Aisenbrey, E.A. and W.L. Murphy, Synthetic alternatives to Matrigel. Nature Reviews Materials, 2020. 5(7): p. 539-551.
28. Carletti, E., A. Motta, and C. Migliaresi, Scaffolds for tissue engineering and 3B cell culture. 3B Cell Culture: Methods and Protocols, 2011: p. 17-39.
29. Hoarau-Véchot, J., et al., Halfway between 2B and animal models: are 3B cultures the ideal tool to study cancer-microenvironment interactions? International journal of molecular sciences, 2018. 19(1): p. 181.
30. Alghuwainem, A., A.T. Alshareeda, and B. Alsowayan, Scaffold-free 3-D cell sheet technique bridges the gap between 2-D cell culture and animal models. International journal of molecular sciences, 2019. 20(19): p. 4926.
31. Timmins, N.E. and L.K. Nielsen, Generation of multicellular tumor spheroids by the hanging-drop method. Tissue engineering, 2007: p. 141-151.
32. Amaral, R.L., et al., Comparative analysis of 3B bladder tumor spheroids obtained by forced floating and hanging drop methods for drug screening. Frontiers in physiology, 2017. 8: p. 605.
33. Barrila, J., et al., Organotypic 3B cell culture models: using the rotating wall vessel to study host–pathogen interactions. Nature Reviews Microbiology, 2010. 8(11): p. 791-801.
34. Basu, A., et al., Ready to go 3B? A semi-automated protocol for microwell spheroid arrays to increase scalability and throughput of 3B cell culture testing. Toxicology Mechanisms and Methods, 2020. 30(8): p. 590- 604.
35. Jorgensen, C. and M. Simon, In vitro human joint models combining advanced 3B cell culture and cutting- edge 3B bioprinting technologies. Cells, 2021. 10(3): p. 596.
36. Russo, M., C.M. Cejas, and G. Pitingolo, Advances in microfluidic 3B cell culture for preclinical drug development. Progress in Molecular Biology and Translational Science, 2022. 187(1): p. 163-204.
37. Li, L., et al., A microfluidic 3B hepatocyte chip for hepatotoxicity testing of nanoparticles. Nanomedicine, 2019. 14(16): p. 2209-2226.
38. Van Duinen, V., et al., Microfluidic 3B cell culture: from tools to tissue models. Current opinion in biotechnology, 2015. 35: p. 118-126.
39. Caleffi, J.T., et al., Magnetic 3B cell culture: State of the art and current advances. Life Sciences, 2021. 286: p. 120028.
40. Anil‐Inevi, M., et al., Magnetic levitation assisted biofabrication, culture, and manipulation of 3B cellular structures using a ring magnet based setup. Biotechnology and Bioengineering, 2021. 118(12): p. 4771-4785.
41. Bonnier, F., et al., Cell viability assessment using the Alamar blue assay: a comparison of 2B and 3B cell culture models. Toxicology in vitro, 2015. 29(1): p. 124-131.
42. Soares, C.P., et al., 2B and 3B-organized cardiac cells shows differences in cellular morphology, adhesion junctions, presence of myofibrils and protein expression. PloS one, 2012. 7(5): p. e38147.
43. Rodríguez-Hernández, M.A., et al., Differential effectiveness of tyrosine kinase inhibitors in 2B/3B culture according to cell differentiation, p53 status and mitochondrial respiration in liver cancer cells. Cell Death & Disease, 2020. 11(5): p. 339.
44. Luca, A.C., et al., Impact of the 3B microenvironment on phenotype, gene expression, and EGFR inhibition of colorectal cancer cell lines. PloS one, 2013. 8(3): p. e59689.
45. Liu, H., J. Lin, and K. Roy, Effect of 3B scaffold and dynamic culture condition on the global gene expression profile of mouse embryonic stem cells. Biomaterials, 2006. 27(36): p. 5978-5989.
46. Baharvand, H., et al., Differentiation of human embryonic stem cells into hepatocytes in 2B and 3B culture systems in vitro. International journal of developmental biology, 2004. 50(7): p. 645-652.
47. Padmalayam, I. and M.J. Suto, 3B cell cultures: Mimicking in vivo tissues for improved predictability in drug discovery. Annual reports in medicinal chemistry, 2012. 47: p. 367-378.
48. Breslin, S. and L. O'Driscoll, The relevance of using 3B cell cultures, in addition to 2B monolayer cultures, when evaluating breast cancer drug sensitivity and resistance. Oncotarget, 2016. 7(29): p. 45745-45756.
49. Souza, A.G., et al., Comparative assay of 2B and 3B cell culture models: proliferation, gene expression and anticancer drug response. Current pharmaceutical design, 2018. 24(15): p. 1689-1694.
50. Imamura, Y., et al., Comparison of 2B-and 3B-culture models as drug-testing platforms in breast cancer. Oncology reports, 2015. 33(4): p. 1837-1843.
51. Melissaridou, S., et al., The effect of 2B and 3B cell cultures on treatment response, EMT profile and stem cell features in head and neck cancer. Cancer cell international, 2019. 19(1): p. 1-10.
52. Wendt, D., et al., Potential and bottlenecks of bioreactors in 3B cell culture and tissue manufacturing. Advanced materials, 2009. 21(32‐33): p. 3352-3367.

Toplam 52 adet kaynakça vardır.

Ayrıntılar

Birincil Dil	Türkçe
Konular	Sinirbilim (Diğer)
Bölüm	Derleme
Yazarlar	Güneş Kibar 0000-0002-2586-6770 Atakan Tevlek 0000-0003-0601-8642
Proje Numarası	123M943
Yayımlanma Tarihi	30 Haziran 2025
Gönderilme Tarihi	25 Aralık 2024
Kabul Tarihi	4 Haziran 2025
Yayımlandığı Sayı	Yıl 2025 Cilt: 34 Sayı: 2

Kaynak Göster

AMA	Kibar G, Tevlek A. İki-boyutlu ve Üç-boyutlu Hücre Kültür Modellerinde Güncel Gelişmeler ve Uygulamalar. aktd. Haziran 2025;34(2):119-128. doi:10.17827/aktd.1607163

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