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1- Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2- Applied Cell Sciences Division, Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran ,s_vahdat@modares.ac.ir
2- Applied Cell Sciences Division, Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran ,
Abstract: (257 Views)
Background and Objectives: It has been demonstrated that three-dimensional (3D) interactions of myeloma cells with other cells and components of the extracellular matrix (ECM) in the bone marrow (BM) microenvironment play critical roles in the disease progression. Therefore, in vitro fabrication of interactions would advance the fields of disease modeling and drug screening. In this regard, we tried to develop simple, cost-beneficial, and available 3D culture conditions for U266 cells using co-culture systems in order to be closer to their native microenvironment.
Materials and Methods: U266 cells, BM mesenchymal stem cells (BM-MSCs) and human umbilical vein endothelial cells (HUVECs) were co-cultured in different matrix-based and matrix-free conditions, and thereafter, the survival and proliferation rates of U266 cells were evaluated and compared. Peripheral blood (PB) plasma was used to generate fibrin gels for matrix-based structures.
Results: Co-cultured cells in the gel-free group could generate and assemble into 3D cell structures. Moreover, the viability and expansion fold of U266 cells in the gel-free group were significantly higher compared to the on-gel and inside-gel groups.
Conclusion: Our simple gel-free 3D co-culture of U266 cells with BM-MSCs and HUVECs, which showed higher viability and proliferation rates of myeloma cells, can be used for further in vitro studies, including drug screening.
Materials and Methods: U266 cells, BM mesenchymal stem cells (BM-MSCs) and human umbilical vein endothelial cells (HUVECs) were co-cultured in different matrix-based and matrix-free conditions, and thereafter, the survival and proliferation rates of U266 cells were evaluated and compared. Peripheral blood (PB) plasma was used to generate fibrin gels for matrix-based structures.
Results: Co-cultured cells in the gel-free group could generate and assemble into 3D cell structures. Moreover, the viability and expansion fold of U266 cells in the gel-free group were significantly higher compared to the on-gel and inside-gel groups.
Conclusion: Our simple gel-free 3D co-culture of U266 cells with BM-MSCs and HUVECs, which showed higher viability and proliferation rates of myeloma cells, can be used for further in vitro studies, including drug screening.
Type of Study: Original Research Article |
Subject:
Medical Biology
Received: 2025/08/13 | Accepted: 2026/04/12
Received: 2025/08/13 | Accepted: 2026/04/12
References
1. Malard F, Neri P, Bahlis NJ, Terpos E, Moukalled N, Hungria VTM, et al. Multiple myeloma. Nat Rev Dis Primers. 2024;10(1):45. [DOI:10.1038/s41572-024-00529-7] [PMID]
2. Mondello P, Cuzzocrea S, Navarra M, Mian M. Bone marrow micro-environment is a crucial player for myelomagenesis and disease progression. Oncotarget. 2017;8(12):20394. [DOI:10.18632/oncotarget.14610] [PMID] [PMCID]
3. Papadimitriou K, Kostopoulos IV, Tsopanidou A, Orologas-Stavrou N, Kastritis E, Tsitsilonis O, et al. Ex vivo models simulating the bone marrow environment and predicting response to therapy in multiple myeloma. Cancers (Basel). 2020;12(8):2006. [DOI:10.3390/cancers12082006] [PMID] [PMCID]
4. Ho M, Goh CY, Patel A, Staunton S, O'Connor R, Godeau M, et al. Role of the bone marrow milieu in multiple myeloma progression and therapeutic resistance. Clin Lymphoma Myeloma Leuk. 2020;20(10):e752-e68. [DOI:10.1016/j.clml.2020.05.026] [PMID]
5. Teramachi J, Miki H, Nakamura S, Hiasa M, Harada T, Abe M. Myeloma bone disease: pathogenesis and management in the era of new anti-myeloma agents. J Bone Mineral Metab. 2023;41(3):388-403. [DOI:10.1007/s00774-023-01403-4] [PMID] [PMCID]
6. Ribatti D, Vacca A. New insights in anti-angiogenesis in multiple myeloma. Int J Molec Sci. 2018;19(7):2031. [DOI:10.3390/ijms19072031] [PMID] [PMCID]
7. Giannakoulas N, Ntanasis-Stathopoulos I, Terpos E. The role of marrow microenvironment in the growth and development of malignant plasma cells in multiple myeloma. Int J Molec Sci. 2021;22(9):4462. [DOI:10.3390/ijms22094462] [PMID] [PMCID]
8. Martini S, Drzeniek NM, Stark R, Kollert MR, Du W, Reinke S, et al. Long-term in vitro maintenance of plasma cells in a hydrogel-enclosed human bone marrow microphysiological 3D model system. Biofabrication. 2024;16(4):045005. [DOI:10.1088/1758-5090/ad5dfe] [PMID]
9. Mehdi SH, Nafees S, Mehdi SJ, Morris CA, Mashouri L, Yoon D. Animal models of multiple myeloma bone disease. Front Genet. 2021;12:640954. [DOI:10.3389/fgene.2021.640954] [PMID] [PMCID]
10. Ferrarini M, Marcatti M, Ciceri F, Ferrero E. 3D Models of surrogate multiple myeloma bone marrow microenvironments: Insights on disease pathophysiology and patient-specific response to drugs. Multiple Myeloma. 2021:93. [DOI:10.5772/intechopen.95333]
11. Verbruggen SW, Freeman CL, Freeman FE. Utilizing 3D models to unravel the dynamics of myeloma plasma cells' escape from the bone marrow microenvironment. Cancers. 2024;16(5):889. [DOI:10.3390/cancers16050889] [PMID] [PMCID]
12. Calar K, Plesselova S, Bhattacharya S, Jorgensen M, de la Puente P. Human plasma-derived 3D cultures model breast cancer treatment responses and predict clinically effective drug treatment concentrations. Cancers (Basel). 2020;12(7):1722. [DOI:10.3390/cancers12071722] [PMID] [PMCID]
13. Alhallak K, de la Puente P, Jeske A, Sun J, Muz B, Rettig MP, et al. 3D tissue engineered plasma cultures support leukemic proliferation and induces drug resistance. Leuk Lymphoma. 2021;62(10):2457-65. [DOI:10.1080/10428194.2021.1919657] [PMID]
14. Barbosa MA, Xavier CP, Pereira RF, Petrikaitė V, Vasconcelos MH. 3D cell culture models as recapitulators of the tumor microenvironment for the screening of anti-cancer drugs. Cancers. 2022;14(1):190. [DOI:10.3390/cancers14010190] [PMID] [PMCID]
15. Ferreira L, Gaspar V, Mano J. Design of spherically structured 3D in vitro tumor models-advances and prospects. Acta Biomaterialia. 2018;75:11-34. [DOI:10.1016/j.actbio.2018.05.034] [PMID] [PMCID]
16. Zlei M, Egert S, Wider D, Ihorst G, Wasch R, Engelhardt M. Characterization of in vitro growth of multiple myeloma cells. Exp Hematol. 2007;35(10):1550-61. [DOI:10.1016/j.exphem.2007.06.016] [PMID]
17. Kaur G, Dufour JM. Cell lines: Valuable tools or useless artifacts. Taylor & Francis; 2012. p. 1-5. [DOI:10.4161/spmg.19885] [PMID] [PMCID]
18. Jomehpour M, Khosravi M, Janfada M, Abroun S, Vahdat S. Establishment of a three-dimensional culture condition for the U266 cell line based on peripheral blood plasma-derived fibrin gels. Cell J (Yakhteh). 2023;25(4):229.
19. de la Puente P, Muz B, Gilson RC, Azab F, Luderer M, King J, et al. 3D tissue-engineered bone marrow as a novel model to study pathophysiology and drug resistance in multiple myeloma. Biomaterials. 2015;73:70-84. [DOI:10.1016/j.biomaterials.2015.09.017] [PMID] [PMCID]
20. Xie AW, Binder BYK, Khalil AS, Schmitt SK, Johnson HJ, Zacharias NA, et al. Controlled self-assembly of stem cell aggregates instructs pluripotency and lineage bias. Sci Rep. 2017;7(1):14070. [DOI:10.1038/s41598-017-14325-9] [PMID] [PMCID]
21. de Janon A, Mantalaris A, Panoskaltsis N. Three-dimensional human bone marrow organoids for the study and application of normal and abnormal hematoimmunopoiesis. J Immunol. 2023;210(7):895-904. [DOI:10.4049/jimmunol.2200836] [PMID] [PMCID]
22. Garcia-Ortiz A, Rodriguez-Garcia Y, Encinas J, Maroto-Martin E, Castellano E, Teixido J, et al. The role of tumor microenvironment in multiple myeloma development and progression. Cancers. 2021;13(2). [DOI:10.3390/cancers13020217] [PMID] [PMCID]
23. Huang YH, Almowaled M, Li J, Venner C, Sandhu I, Peters A, et al. Three-dimensional reconstructed bone marrow matrix culture improves the viability of primary myeloma cells in-vitro via a STAT3-dependent mechanism. Curr Issues Mol Biol. 2021;43(1):313-23. [DOI:10.3390/cimb43010026] [PMID] [PMCID]
24. Garcia-Ortiz A, Rodríguez-García Y, Encinas J, Maroto-Martín E, Castellano E, Teixidó J, et al. The role of tumor microenvironment in multiple myeloma development and progression. Cancers. 2021;13(2):217. [DOI:10.3390/cancers13020217] [PMID] [PMCID]
25. Lu K, Wang W, Liu Y, Xie C, Liu J, Xing L. Advancements in microenvironment-based therapies: transforming the landscape of multiple myeloma treatment. Front Oncol. 2024;14:1413494. [DOI:10.3389/fonc.2024.1413494] [PMID] [PMCID]
26. Garayoa M, Garcia JL, Santamaría C, García-Gómez A, Blanco JF, Pandiella A, et al. Mesenchymal stem cells from multiple myeloma patients display distinct genomic profile as compared with those from normal donors. Leukemia. 2009;23(8):1515-27. [DOI:10.1038/leu.2009.65] [PMID]
27. Attar-Schneider O, Zismanov V, Dabbah M, Tartakover-Matalon S, Drucker L, Lishner M. Multiple myeloma and bone marrow mesenchymal stem cells' crosstalk: Effect on translation initiation. Molec Carcinogen. 2016;55(9):1343-54. [DOI:10.1002/mc.22378] [PMID]
28. Melaccio A, Reale A, Saltarella I, Desantis V, Lamanuzzi A, Cicco S, et al. Pathways of angiogenic and inflammatory cytokines in multiple myeloma: role in plasma cell clonal expansion and drug resistance. J Clin Med. 2022;11(21):6491. [DOI:10.3390/jcm11216491] [PMID] [PMCID]
29. Belloni D, Heltai S, Ponzoni M, Villa A, Vergani B, Pecciarini L, et al. Modeling multiple myeloma-bone marrow interactions and response to drugs in a 3D surrogate microenvironment. Haematologica. 2018;103(4):707-16. [DOI:10.3324/haematol.2017.167486] [PMID] [PMCID]
30. Braham MVJ, Minnema MC, Aarts T, Sebestyen Z, Straetemans T, Vyborova A, et al. Cellular immunotherapy on primary multiple myeloma expanded in a 3D bone marrow niche model. Oncoimmunol. 2018;7(6):e1434465. [DOI:10.1080/2162402X.2018.1434465] [PMID] [PMCID]
31. Takebe T, Enomura M, Yoshizawa E, Kimura M, Koike H, Ueno Y, et al. Vascularized and complex organ buds from diverse tissues via mesenchymal cell-driven condensation. Cell Stem Cell. 2015;16(5):556-65. [DOI:10.1016/j.stem.2015.03.004] [PMID]
32. Djomehri SI, Burman B, Gonzalez ME, Takayama S, Kleer CG. A reproducible scaffold-free 3D organoid model to study neoplastic progression in breast cancer. J Cell Commun Signal. 2019;13(1):129-43. [DOI:10.1007/s12079-018-0498-7] [PMID] [PMCID]
33. Varzideh F, Pahlavan S, Ansari H, Halvaei M, Kostin S, Feiz MS, et al. Human cardiomyocytes undergo enhanced maturation in embryonic stem cell-derived organoid transplants. Biomaterials. 2019;192:537-50. [DOI:10.1016/j.biomaterials.2018.11.033] [PMID]
34. Takebe T, Zhang RR, Koike H, Kimura M, Yoshizawa E, Enomura M, et al. Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant. Nat Protoc. 2014;9(2):396-409. [DOI:10.1038/nprot.2014.020] [PMID]
35. Li Y, Meng H, Liu Y, Lee BP. Fibrin gel as an injectable biodegradable scaffold and cell carrier for tissue engineering. Sci W J. 2015;2015. [DOI:10.1155/2015/685690] [PMID] [PMCID]
36. Shoji T, Nakasa T, Yoshizuka M, Yamasaki T, Yasunaga Y, Adachi N, et al. Comparison of fibrin clots derived from peripheral blood and bone marrow. Connect Tissue Res. 2017;58(2):208-14. [DOI:10.1080/03008207.2016.1215443] [PMID]
37. Garcia-Briega MI, Pla-Salom J, Clara-Trujillo S, Tolosa L, Cordon L, Sempere A, et al. Co-culture of multiple myeloma cell lines and bone marrow mesenchymal stem cells in a 3D microgel environment. Biomaterials advances. 2025;172:214243. [DOI:10.1016/j.bioadv.2025.214243] [PMID]
38. Heo DN, Hospodiuk M, Ozbolat IT. Synergistic interplay between human MSCs and HUVECs in 3D spheroids laden in collagen/fibrin hydrogels for bone tissue engineering. Acta biomaterialia. 2019;95:348-56. [DOI:10.1016/j.actbio.2019.02.046] [PMID]
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