| تعداد نشریات | 24 |
| تعداد شمارهها | 929 |
| تعداد مقالات | 8,346 |
| تعداد مشاهده مقاله | 16,803,032 |
| تعداد دریافت فایل اصل مقاله | 14,117,375 |
Cloning and expressing of interleukine 2 in amniotic membrane-derived mesenchymal stem cells, as a potent feeder layer | ||
| Molecular Biology Research Communications | ||
| دوره 10، شماره 2، 2021، صفحه 63-71 اصل مقاله (528.17 K) | ||
| نوع مقاله: Original article | ||
| شناسه دیجیتال (DOI): 10.22099/mbrc.2021.38845.1566 | ||
| نویسندگان | ||
| Saeid Anvari1، 2؛ Farshad Foroughi3؛ Mehdi Azad4؛ Amirhosein Maali1، 5؛ Safar Ali Alizadeh6؛ Mohammad Hossein Ahmadi* 4 | ||
| 1Department of Medical Biotechnology, Faculty of Allied Medicine, Qazvin University of Medical Sciences, Qazvin, Iran | ||
| 2Student Research Committee, Qazvin University of Medical Sciences, Qazvin, Iran | ||
| 3Department of Immunology, School of Medicine, Qazvin University of Medical Sciences | ||
| 4Department of Medical Laboratory Sciences, Faculty of Allied Medicine, Qazvin University of Medical Sciences, Qazvin, Iran | ||
| 5Student Research Committee, Pasteur Institute of Iran, Tehran, Iran | ||
| 6Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran | ||
| چکیده | ||
| The application of mesenchymal stem cells (MSCs) is rapidly expanding due to their unique properties in cell therapy, especially as the feeder layer in the ex-vivo expansion of immune cells. Also, Interleukin 2 (IL-2) is an essential human cytokine in the expansion of hematopoietic precursors and progenitors, i.e., NK cells and T cells, while there is no endogenous expression of IL-2 in MSCs. This study aimed to examine the potency of amniotic membrane (AM)-MSCs as the IL-2 secretory cells. IL-2-containingpCMV3-C-GFPspark shuttle vector was transformed in E.coli DH5-alpha. After cloning, the plasmid DNA was extracted and transfected in isolated AM-MSCs, by lipofectamine-2000. Then, the RNA and protein expression levels of exogenous IL-2 were evaluated 3 to 15 days after transfection, using ELISA and qRT-PCR. Fluorescent microscopy and flowcytometry assays were used for evaluating the GFP-positivity of transfected AM-MSCs, as IL-2 expression control. There was a significant increase in RNA expression of exogenous IL-2 in transfected AM-MSCs in 3 to 15 days after transfection. (p <0.001) Also, IL-2 concentration released in the medium was increased in 3rd day after transfection (611 pg/ml). However, the RNA and protein expression of IL-2 was reduced through passing the time. The results show AM-MSC is a suitable host for the expression and secretion of IL-2 as a critical cytokine in the ex-vivo expansion of hematopoietic precursors and progenitors, i.e., NK cells and T cells. Also, the survival time of IL-2 expression in AM-MSCs was long enough for use as a feeder layer. | ||
| کلیدواژهها | ||
| Interleukin-2؛ Mesenchymal stem cells؛ Plasmids؛ Transfection | ||
|
سایر فایل های مرتبط با مقاله
|
||
| مراجع | ||
|
1. Bondanza A, Bonini C, Fehse B, Hudecek M. Cellular Therapy with Engineered T Cells, Efficacy and Side Effects. In: Carreras E, Dufour C, Mohty M, Kröger N, editors. The EBMT Handbook: Hematopoietic Stem Cell Transplantation and Cellular Therapies. Cham (CH): Springer Copyright 2019, EBMT and the Author(s). 2019. p. 449-56.
2. Hayes C. Cellular immunotherapies for cancer. Ir J Med Sci 2021;190:41-57.
3.Wiltrout RH. Regulation and antimetastatic functions of liver-associated natural killer cells. Immunol Rev 2000;174:63-76.
4. Biron CA, Brossay L. NK cells and NKT cells in innate defense against viral infections. Curr Opin Immunol 2001;13:458-464.
5. Song Y, Hu B, Liu Y, Jin Z, Zhang Y, Lin D, Zhu Y, Lei L, Gong H, Mei Y, Teo HY, Wu D, Liu H. IL-12/IL-18-preactivated donor NK cells enhance GVL effects and mitigate GvHD after allogeneic hematopoietic stem cell transplantation. Eur J Immunol 2018;48:670-682.
6. Moradi Z, Maali A, Shad JS, Farasat A, Kouchaki R, Moghadami M, Ahmadi MH, Azad M. Updates on novel erythropoiesis-stimulating agents: Clinical and molecular approach. Indian J Hematol Blood Transfus 2020;36:26-36.
7. Ghorban K, Shanaki M, Mobarra N, Azad M, Asadi J, Pakzad R, Ehteram H. Apolipoproteins A1, B, and other prognostic biochemical cardiovascular risk factors in patients with beta-thalassemia major. Hematology 2016;21:113-120.
8. Maroufi F, Maali A, Abdollahpour-Alitappeh M, Ahmadi MH, Azad M. CRISPR-mediated modification of DNA methylation pattern in the new era of cancer therapy. Epigenomics 2020;12:1845-1859.
9. Bald T, Krummel MF, Smyth MJ, Barry KC. The NK cell-cancer cycle: advances and new challenges in NK cell-based immunotherapies. Nat Immunol 2020;21:835-847.
10. Arai S, Meagher R, Swearingen M, Myint H, Rich E, Martinson J, Klingemann H. Infusion of the allogeneic cell line NK-92 in patients with advanced renal cell cancer or melanoma: a phase I trial. Cytotherapy 2008;10:625-632.
11. Iliopoulou EG, Kountourakis P, Karamouzis MV, Doufexis D, Ardavanis A, Baxevanis CN, Rigatos G, Papamichail M, Perez SA. A phase I trial of adoptive transfer of allogeneic natural killer cells in patients with advanced non-small cell lung cancer. Cancer Immunol Immunother 2010;59:1781-1789.
12. Geller MA, Miller JS. Use of allogeneic NK cells for cancer immunotherapy. Immunotherapy 2011;3:1445-1459.
13. Geller MA, Cooley S, Judson PL, Ghebre R, Carson LF, Argenta PA, Jonson AL, Panoskaltsis-Mortari A, Curtsinger J, McKenna D, Dusenbery K, Bliss R, Downs LS, Miller JS. A phase II study of allogeneic natural killer cell therapy to treat patients with recurrent ovarian and breast cancer. Cytotherapy 2011;13:98-107.
14. Levy EM, Roberti MP, Mordoh J. Natural killer cells in human cancer: from biological functions to clinical applications. J Biomed Biotechnol 2011;2011:676198.
15. Boissel L, Tuncer HH, Betancur M, Wolfberg A, Klingemann H. Umbilical cord mesenchymal stem cells increase expansion of cord blood natural killer cells. Biol Blood Marrow Transplant 2008;14:1031-1038.
16. Meng X, Sun B, Xiao Z. Comparison in transcriptome and cytokine profiles of mesenchymal stem cells from human umbilical cord and cord blood. Gene 2019;696:10-20.
17. Lai RC, Yeo RW, Lim SK. Mesenchymal stem cell exosomes. Semin Cell Dev Biol 2015;40:82-88.
18. Han Y, Li X, Zhang Y, Han Y, Chang F, Ding J. Mesenchymal Stem Cells for Regenerative Medicine. Cells 2019;8:886.
19. Fu X, Liu G, Halim A, Ju Y, Luo Q, Song AG. Mesenchymal Stem Cell Migration and Tissue Repair. Cells 2019;8:784.
20. Roubelakis MG, Trohatou O, Anagnou NP. Amniotic fluid and amniotic membrane stem cells: marker discovery. Stem Cells Int 2012;2012:107836.
21. Rautela J, Huntington ND. IL-15 signaling in NK cell cancer immunotherapy. Curr Opin Immunol 2017;44:1-6.
22. Cifaldi L, Pinto RM, Rana I, Caniglia M, Angioni A, Petrocchi S, Cancrini C, Cursi L, Palumbo G, Zingoni A, Gismondi A, Rossi P, Santoni A, Cerboni C. NK cell effector functions in a Chediak-Higashi patient undergoing cord blood transplantation: Effects of in vitro treatment with IL-2. Immunol Lett 2016;180:46-53.
23. Masuyama J. NK Cell Culture Container And NK Cell Culture Method. Google Patents; 2017.
24. Ribeiro S, Mairhofer J, Madeira C, Diogo MM, Lobato da Silva C, Monteiro G, Grabherr R, Cabral JM. Plasmid DNA size does affect nonviral gene delivery efficiency in stem cells. Cell Reprogram. 2012;14:130-137.
25.Srinivasan C, Burgess DJ. Optimization and characterization of anionic lipoplexes for gene delivery. J Control Release 2009;136:62-70.
26. Madeira C, Mendes RD, Ribeiro SC, Boura JS, Aires-Barros MR, da Silva CL, Cabral JM. Nonviral gene delivery to mesenchymal stem cells using cationic liposomes for gene and cell therapy. J Biomed Biotechnol 2010;2010:735349.
27. Cohen RN, van der Aa MA, Macaraeg N, Lee AP, Szoka FC, Jr. Quantification of plasmid DNA copies in the nucleus after lipoplex and polyplex transfection. J Control Release 2009; 135:166-174.
28. Kamiya H, Fujimura Y, Matsuoka I, Harashima H. Visualization of intracellular trafficking of exogenous DNA delivered by cationic liposomes. Biochem Biophys Res Commun 2002; 298:591-597.
29. Gheisari Y, Soleimani M, Azadmanesh K, Zeinali S. Multipotent mesenchymal stromal cells: optimization and comparison of five cationic polymer-based gene delivery methods. Cytotherapy 2008;10:815-823.
30. Helledie T, Nurcombe V, Cool SM. A simple and reliable electroporation method for human bone marrow mesenchymal stem cells. Stem Cells Dev 2008;17:837-848.
31. Nagai K, Harada Y, Harada H, Yanagihara K, Yonemitsu Y, Miyazaki Y. Highly Activated Ex Vivo-expanded Natural Killer Cells in Patients With Solid Tumors in a Phase I/IIa Clinical Study. Anticancer Res 2020;40:5687-5700. | ||
|
آمار تعداد مشاهده مقاله: 2,410 تعداد دریافت فایل اصل مقاله: 1,246 |
||
