Volume 22, Issue 5 (Iranian South Medical Journal 2019)
Iran South Med J 2019, 22(5): 278-295 |
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Vazifeh shiran N, Abroun S. Immunophenotypic and Immunotypic Evalua-tions of Human Myeloma Cell Lines KMM-1, JJN3, LP1, L363, KMS-12BM and RPMI-8226 for Cell-Line Authentication. Iran South Med J 2019; 22 (5) :278-295
URL: http://ismj.bpums.ac.ir/article-1-1172-en.html
URL: http://ismj.bpums.ac.ir/article-1-1172-en.html
1- Department. of Hematology, School of Medical Sciences, Tarbiate Modares University, Tehran, Iran , nadershirani@yahoo.com
2- Department. of Hematology, School of Medical Sciences, Tarbiate Modares University, Tehran, Iran
2- Department. of Hematology, School of Medical Sciences, Tarbiate Modares University, Tehran, Iran
Abstract: (3845 Views)
Background: The disease-associated cell lines used in the majority of preclinical trials should be first authenticated and identified to avoid directing research expenditure on wrong cells and obtaining irrelevant results. The present study evaluated the immunophenotypic and immunotypic authenticity of six human myeloma cell lines (HMCLs).
Materials and Methods: Cytospin smear and Wright staining were used for invetigating cell morphology, the flowcytometry of the markers CD45, CD2, CD19, CD38 and CD138 for immunophenotypic investigations, capillary electrophoresis for the immunoglobulin secretion potency and immunofixation for the immunotypical investigation of the cells.
Results: Given the definite plasma cell morphology, the identity of all the cell lines was confirmed, although a similar morphology was observed in L363 and JJN3. In addition to authenticating the myeloma nature of all the cell lines, immunophenotypic investigations suggested a complete similarity between L363 and JJN3 cell lines. Examining their immunotypes for a final confirmation found KMS12BM to be non-secretor, KMM1 to be BJP-λ and the remaining to be IgG-λ.
Conclusion: Despite confirming the identity of four cell lines, i.e. RPMI-8226, KMS-12BM, KMM1 and LP1, that of JJN3 and L363 cells, whose immunotype was determined as IgGλ, was questioned owing to their fundamental morphological, immunophenotypic and immunotypic similarities. On the other hand, a reveiew of literature and ATCC.org suggested that the main nature of JJN3 is IgAκ. The authenticity of the JJN3 cell line was therefore unclear and it was indeed the same as that of L363. The JJN3 cell-line was therefore discarded, and the associated results were interpreted as consistent with the results of L363.
Materials and Methods: Cytospin smear and Wright staining were used for invetigating cell morphology, the flowcytometry of the markers CD45, CD2, CD19, CD38 and CD138 for immunophenotypic investigations, capillary electrophoresis for the immunoglobulin secretion potency and immunofixation for the immunotypical investigation of the cells.
Results: Given the definite plasma cell morphology, the identity of all the cell lines was confirmed, although a similar morphology was observed in L363 and JJN3. In addition to authenticating the myeloma nature of all the cell lines, immunophenotypic investigations suggested a complete similarity between L363 and JJN3 cell lines. Examining their immunotypes for a final confirmation found KMS12BM to be non-secretor, KMM1 to be BJP-λ and the remaining to be IgG-λ.
Conclusion: Despite confirming the identity of four cell lines, i.e. RPMI-8226, KMS-12BM, KMM1 and LP1, that of JJN3 and L363 cells, whose immunotype was determined as IgGλ, was questioned owing to their fundamental morphological, immunophenotypic and immunotypic similarities. On the other hand, a reveiew of literature and ATCC.org suggested that the main nature of JJN3 is IgAκ. The authenticity of the JJN3 cell line was therefore unclear and it was indeed the same as that of L363. The JJN3 cell-line was therefore discarded, and the associated results were interpreted as consistent with the results of L363.
Type of Study: Original |
Subject:
Hemic and Lymphatic Systems
Received: 2019/07/7 | Accepted: 2019/08/21 | Published: 2019/12/1
Received: 2019/07/7 | Accepted: 2019/08/21 | Published: 2019/12/1
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9. Flores-Montero J, De Tute R, Paiva B, et al. Immunophenotype of Normal vs. Myeloma Plasma Cells: Toward Antibody Panel Specifications for MRD Detection in Multiple Myeloma. Cytometry B Clin Cytom 2016; 90(1): 61-72. [DOI:10.1002/cyto.b.21265]
10. Kawano Y, Fujiwara S, Wada N, et al. Multiple Myeloma Cells Expressing Low Levels of CD138 Have an Immature Phenotype and Reduced Sensitivity to Lenalidomide. Int J Oncol 2012; 41(3): 876-84. [DOI:10.3892/ijo.2012.1545]
11. Togawa A, Inoue N, Miyamoto K, et al. Establishment and Characterization of a Human Myeloma Cell Line (KMM-1). Int J Cancer 1982; 29(5): 495-500. [DOI:10.1002/ijc.2910290502]
12. Moreaux J, Klein B, Bataille R, et al. A High-Risk Signature for Patients with Multiple Myeloma Established from the Molecular Classification of Human Myeloma Cell Lines. Haematologica 2011; 96(4): 574-82. [DOI:10.3324/haematol.2010.033456]
13. Vincent Rajkumar S. Multiple Myeloma: 2018 Update on Diagnosis, Risk‐Stratification, and Management. Am J Hematol 2018; 93(8): 1091-110. [DOI:10.1002/ajh.25117]
14. Drexler HG, Matsuo Y. Malignant Hematopoietic Cell Lines: in Vitro Models for the Study of Multiple Myeloma and Plasma Cell Leukemia. Leuk Res 2000; 24(8): 681-703. [DOI:10.1016/S0145-2126(99)00195-2]
15. Ohtsuki T, Yawata Y, Wada H, et al. Two Human Myeloma Cell Lines, Amylase-Producing KMS-12-PE and Amylase-Non-Producing KMS-12-BM, Were Established from a Patient, Having the Same Chromosome Marker, t(l1;14)(ql3;q32). Br J Haematol 1989; 73(2): 199-204. [DOI:10.1111/j.1365-2141.1989.tb00252.x]
16. Klein U, Dalla-Favera R. Germinal Centres: Role in B-Cell Physiology and Malignancy. Nat Rev Immunol 2008; 8(1): 22-33. [DOI:10.1038/nri2217]
17. Nakaya A, Fujita S, Satake A, et al. Impact of CRAB Symptoms in Survival of Patients with Symptomatic Myeloma in Novel Agent Era. Hematol Rep 2017; 9(1): 6887. [DOI:10.4081/hr.2017.6887]
18. Nutt SL, Hodgkin PD, Tarlinton DM, et al. The Generation of Antibody-Secreting Plasma Cells. Nat Rev Immunol 2015; 15(3): 160-71. [DOI:10.1038/nri3795]
19. Zhan F, Huang Y, Colla S, et al. The Molecular Classification of Multiple Myeloma. Blood 2006; 108(6): 2020-28. [DOI:10.1182/blood-2005-11-013458]
20. Manzanera MG, Izquierdo SJ, Matos OA. Immunophenotyping of Plasma Cells in Multiple Myeloma. Methods Mol Med 2005; 113: 5-24. [DOI:10.1385/1-59259-916-8:5]
21. Gooding PR, Bybee A, Cooke A, et al. Phenotypic and Molecular Analysis of Six Human Cell Lines Derived from Patients with Plasma Cell Dyscrasia. Br J Haematol 1999; 106(3): 669-81. [DOI:10.1046/j.1365-2141.1999.01602.x]
22. Bakhtiari M, Kaviani S, Abroon S, Moallemi M, Jalaei Kho H. Amniotic Fluid that Supports the Growth of Plasma Cells. Iran South Med J 2018; 21(4): 304-318.
23. Seegmiller AC, Xu Y, McKenna RW, et al. Immunophenotypic Differentiation between Neoplastic Plasma Cells in Mature B-cell lymphoma vs Plasma Cell Myeloma. Am J Clin Pathol 2007; 127(2): 176-81. [DOI:10.1309/5EL22BH45PHUPM8P]
24. Flores-Montero J, De Tute R, Paiva B, et al. Immunophenotype of Normal vs. Myeloma Plasma Cells: Toward Antibody Panel Specifications for MRD Detection in Multiple Myeloma. Cytometry B Clin Cytom 2016; 90(1): 61-72. [DOI:10.1002/cyto.b.21265]
25. Kawano Y, Fujiwara S, Wada N, et al. Multiple Myeloma Cells Expressing Low Levels of CD138 Have an Immature Phenotype and Reduced Sensitivity to Lenalidomide. Int J Oncol 2012; 41(3): 876-84. [DOI:10.3892/ijo.2012.1545]
26. Togawa A, Inoue N, Miyamoto K, et al. Establishment and Characterization of a Human Myeloma Cell Line (KMM-1). Int J Cancer 1982; 29(5): 495-500. [DOI:10.1002/ijc.2910290502]
27. Moreaux J, Klein B, Bataille R, et al. A High-Risk Signature for Patients with Multiple Myeloma Established from the Molecular Classification of Human Myeloma Cell Lines. Haematologica 2011; 96(4): 574-82. [DOI:10.3324/haematol.2010.033456]
28. Vincent Rajkumar S. Multiple Myeloma: 2018 Update on Diagnosis, Risk‐Stratification, and Management. Am J Hematol 2018; 93(8): 1091-110. [DOI:10.1002/ajh.25117]
29. Drexler HG, Matsuo Y. Malignant Hematopoietic Cell Lines: in Vitro Models for the Study of Multiple Myeloma and Plasma Cell Leukemia. Leuk Res 2000; 24(8): 681-703. [DOI:10.1016/S0145-2126(99)00195-2]
30. Ohtsuki T, Yawata Y, Wada H, et al. Two Human Myeloma Cell Lines, Amylase-Producing KMS-12-PE and Amylase-Non-Producing KMS-12-BM, Were Established from a Patient, Having the Same Chromosome Marker, t(l1;14)(ql3;q32). Br J Haematol 1989; 73(2): 199-204. [DOI:10.1111/j.1365-2141.1989.tb00252.x]
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